To see the other types of publications on this topic, follow the link: Lipid transfer proteins.
Dissertations / Theses on the topic 'Lipid transfer proteins'
Create a spot-on reference in APA, MLA, Chicago, Harvard, and other styles
Consult the top 50 dissertations / theses for your research on the topic 'Lipid transfer proteins.'
Next to every source in the list of references, there is an 'Add to bibliography' button. Press on it, and we will generate automatically the bibliographic reference to the chosen work in the citation style you need: APA, MLA, Harvard, Chicago, Vancouver, etc.
You can also download the full text of the academic publication as pdf and read online its abstract whenever available in the metadata.
Browse dissertations / theses on a wide variety of disciplines and organise your bibliography correctly.
1
Edstam, Monika. "Plant lipid transfer proteins : Evolution, expression and function." Doctoral thesis, Linköpings universitet, Biologi, 2013. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-98117.
Full textAbstract:
The plant non-specific lipid transfer proteins (nsLTPs) are known for the ability to transfer different lipids in vitro, but their in vivo functions have not yet been elucidated. They seem to play a role in the defense against biotic and abiotic stresses; the gene expression of nsLTPs is often upregulated when exposed to stresses. Further, two different nsLTPs have been shown to affect the lipid composition of the plant cuticle, a structure acting as a protective barrier. However, more evidence is needed to prove this hypothesis and to pinpoint their exact role in this process. In this thesis I have shown that the nsLTPs are found in all land plants, but not in any of the studied algae. This supports a role in defense response, since protection against dehydration, radiation, pathogens and other stresses played a crucial role when plants adapted to a life on land. Characterization of the nsLTPs in early diverging land plant revealed that even though the amino acid similarity towards nsLTPs in flowering plants is not very high, the main properties of the proteins are still the same (Paper I). This includes the protein structure, which consists of α-helices surrounding a lipid binding cavity, a conserved pattern of cysteine residues involved in disulphide bonds and a signal sequence directing the protein to the extracellular space. Further, the expression of nsLTPs in the moss Physcomitrella patens was shown to respond to stresses, and construction of an YFP-LTP fusion protein confirmed the localization to the periphery of the cell in planta (Paper II). Heterologous expressed Physcomitrella nsLTPs were also shown to have the ability to bind lipids and to be very heat stable, features previously only studied in nsLTPs from flowering plants. By examining the presence of a cuticle in Physcomitrella, a correlation between the nsLTPs´ lipid binding ability and the lipid composition of the cuticle could be found, which further strengthens the involvement of nsLTPs in transfer of lipids for cuticle construction. In the flowering plant Arabidopsis thaliana, I showed that several of the nsLTPs followed the same expression pattern when examining data from different tissues, stress treatments, hormones, chemical treatments and developmental stages, but also that four of the genes were undergoing alternative splicing resulting in different isoforms of the proteins (Paper III). Based on their expression patterns, the genes could be divided into three different coexpression networks. By examining other genes similarly expressed, each network could be designated to a putative function: Transfer of lipids for synthesis of the cuticle, suberin layer and sporopollenin, respectively. In Paper IV, these hypotheses were tested in vivo by examining knockout mutants of several nsLTPs in Arabidopsis. The involvement in sporopollenin deposition could be confirmed; two of the knockout lines showed collapsed pollen grains. Further, two other lines showed an increased seed coat permeability due to an altered lipid composition of the suberin layer. Together, the results support a role for nsLTPs in construction of the protecting barriers in all land plants.
2
Jülke, Sabine. "Lipid-Transfer-Proteine aus Arabidopsis thaliana - physiologische und molekulare Funktionsanalyse." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2013. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-104357.
Full textAbstract:
Die durch den obligat biotrophen Protisten Plasmodiophora brassicae hervorgerufene Pflanzenkrankheit Kohlhernie verursacht weltweit hohe ökonomische Verluste. Bis heute gibt es keine effektiven Möglichkeiten, diese Pflanzenkrankheit zu bekämpfen. Eine Analyse der Genexpression in infizierten Wurzeln im Vergleich zu nicht infizierten Wurzeln ergab, dass die Gene für Lipid-Transfer-Proteine während der gesamten Krankheitsentwicklung differentiell reguliert sind. Über die Funktionen von Lipid-Transfer-Proteinen in Pflanzen wird noch spekuliert. Diskutiert wird dabei eine Funktion bei der Anpassung an verschiedene abiotische Stressfaktoren, bei der Pathogenabwehr sowie bei dem Transfer von Lipiden.
In dieser Arbeit wurden transgene Pflanzen generiert, in denen die pathogenbedingte LTP-Genregulation umgekehrt ist. Es wurden transgene A. thaliana Pflanzen erzeugt, die die Gene LTP1, LTP3, LTP4, AT1G12090 sowie AT2G18370 überexprimieren und die Genexpression von AT4G33550 sowie AT1G62510 reprimieren. Die Regulation der LTP-Genexpression erfolgte dabei durch den wurzel- und keimlingsspezifischen Promotor Pyk10. Zusätzlich wurden in dieser Arbeit auch T-DNA-Insertionsmutanten für die Gene AT1G12090, AT2G18370, AT3G22620, AT5G05960, LTP3 sowie LTP4 untersucht. Mittels semiquantitativer Expressionsanalyse konnte die Modulation der LTP-Genexpression in den LTP-Mutanten bestätigt werden. Darüber hinaus konnte gezeigt werden, dass die Modulation der Expression eines LTP-Gens auch die Expression anderer LTP-Gene beeinflusst.
Die phytopathologischen Analysen der LTP-Mutanten hinsichtlich der Entwicklung der Pflanzenkrankheit Kohlhernie ergab, dass die Überexpression der Gene LTP1, LTP3 sowie AT2G18370 und die Repression der Expression von AT1G62510 eine verringerte Anfälligkeit für diese Krankheit bewirkt. Die verstärkte Expression der Gene LTP1, LTP3, LTP4, AT1G12090 sowie AT2G18370 resultiert außerdem in einer verringerten Symptomentwicklung infolge einer Pseudomonas syringae-Infektion. Die verringerte Expression des Gens AT4G33550 führt hingegen zu einer größeren Anfälligkeit für eine P. brassicae Infektion; die Infektion mit P. syringae wird dadurch aber nicht beeinflusst. Die physiologische Charakterisierung der LTP-Mutanten umfasste die Analyse des Pflanzenwachstums unter Salzstress bzw. osmotischem Stress sowie die Entwicklung der Seneszenz in abgetrennten Rosettenblättern. Es konnte gezeigt werden, dass die Gene LTP1, LTP3, LTP4, AT4G33550 sowie AT1G62510 bei der Anpassung an Salzstress sowie die Gene LTP3, AT3G22620, AT4G33550 und AT1G62510 bei der Anpassung an osmotischen Stress eine Rolle spielen. Durch die Modulation der Expression der genannten Gene wird das Wachstum unter diesen Stressbedingungen sowohl positiv als auch negativ beeinflusst. Die Entwicklung der Seneszenz wird ebenfalls durch eine veränderte LTP-Genexpression (LTP1, LTP3, LTP4, AT3G22620 sowie AT4G33550) beeinflusst.
Für die biochemische Charakterisierung wurden die LTP-Gene aus A. thaliana mit einem Fusionspartner in E. coli exprimiert und die resultierenden Fusionsproteine gereinigt. Diese wurden nach Abspalten des Fusionspartners hinsichtlich ihrer antimikrobiellen Aktivität und auf die Fähigkeit, Calmodulin zu binden, untersucht. Für die gereinigten Lipid-Transfer-Proteine LTP1, LTP3, LTP4, AT2G18370 sowie AT1G62510 konnte unter den bisher getesteten Versuchsbedingungen keine antimikrobielle Aktivität nachgewiesen werden. Für die Proteine LTP1, LTP3 und LTP4 konnte eine calciumunabhängige Calmodulin-Bindung nachgewiesen werden. Die Ergebnisse dieser Versuche ermöglichen keine Aussage bezüglich der genauen Funktion der einzelnen Lipid-Transfer-Proteine, geben aber Hinweise darauf, dass diese bei den entsprechenden Stress-Vorgängen eine Rolle spielen. Welche Funktion sie dabei genau erfüllen, muss in weiterführenden Analysen untersucht werden.
3
Jansson, Sandra. "Regulation of non-specific lipid transfer proteins in abiotically stressed Physcomitrella patens." Thesis, Linköpings universitet, Molekylär genetik, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-69438.
Full textAbstract:
Non-specific lipid transfer proteins is a large and diverse protein family found in plants, with roles in biological systems ranging from long distance signaling to plant pathogen defense. Little is known about the roles of nsLTPs, but recent studies have cast some light on the issue, among other things proposing that they may be involved in the cutice formation on land-living liverworts, mosses and non-seedbearing plants. Increased cuticle formation is thought to be a part of a plants defense system against stress. In this experiment, the expression of nsLTPs type G in the moss Physcomitrella patens was examined by qRT-PCR on cDNA synthesized from already existing mRNA samples from moss under different abiotic stresses. The different stresses were UV-light, salt (ion toxicity), heavy metal, cold drought, plant hormone and osmosis. House-keeping gene P. patens beta-tubuline 1 was used as reference and relative expression analysis was performed. The study showed a general down-regulation of PpLTPg's in the abiotically stressed samples, and the possible coupled regulatory response of PpLTPg3 and PpLTPg5. The results imply that the PpLTPg's in Physcomitrella patens could be connected to biological processes that cease during stress, or that they worl through negative feedback to support plant defense against stress.
4
Höglund, Andrey. "Expression pattern of GPI-anchored non-specific lipid transfer proteins in Physcomitrella patens." Thesis, Linköpings universitet, Molekylär genetik, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-71702.
Full textAbstract:
During the water-to-land transition, that occurred approximately 450 MYA, novel habitats wererevealed to the emerging plants. This terrestrial habitat was a harsh environment compared to theaquatic, with shifting substrate content, irregular supply of water, damaging UV-radiation andrapid fluctuating temperatures. Non-specific lipid transfer proteins (nsLTP) are today only foundin the land living plants and not in the green algae. This suggests that these genes might haveevolved to help the plants cope with the stressful conditions. In this study the expression patternhas been analysed of the nsLTPs in the moss Physcomitrella patens during the possible conditionsthat raised during the water-to-land transition. The moss was exposed to salt, UV-B, drought, copper, cold and osmotic stress. Quantitative real-time PCR was used to analyse the transcription levels. I found that six genes were upregulated during either cold, dehydration or UV-B stress. This suggest that these genes are involved in the plant defense against these abiotic stresse
5
DeBono, Allan. "The role and behavior of Arabidopsis thaliana lipid transfer proteins during cuticular wax deposition." Thesis, University of British Columbia, 2011. http://hdl.handle.net/2429/39381.
Full textAbstract:
The primary aerial surfaces of terrestrial plants are coated with a protective hydrophobic layer
comprising insoluble and soluble lipids. The lipids are known collectively as cuticular wax. To
generate the waxy cuticle during elongative growth, plants dedicate half of the fatty acid
metabolism of their epidermal cells. It is unknown how cuticular wax is exported from the plasma membrane into the cell wall, and eventually, to the cuticle at the cell surface. I hypothesized that lipid transfer proteins (LTPs) were responsible for plasma membrane to cell wall transport of cuticular lipids. Using an epidermis-specific microarray, I identified five candidate Arabidopsis LTPs. I discovered that mutations in gene At1g27950 result in a stem wax phenotype: reduced cuticular lipid nonacosane resulting in reduced total wax compared to wildtype. This gene encodes a glycosylphosphatidylinositol (GPI)-linked LTP and thus was named
LTPG. In contrast, to LTPG, no detectable wax phenotype was found in mutants for classical LTPs. In
phylogenetic analyses, these LTPs clustered into a weakly related group that I named LTPAs. In
an attempt to overcome genetic redundancy I made double and triple mutants from the candidate
LTPAs. None of these mutants displayed detectable changes in wax compared with wildtype.
Using live cell imaging, I showed that LTPG is localized to the epidermal cell plasma membrane
and the cell wall and accumulates non-uniformly on the plant surface. I employed fluorescence
recovery after photobleaching to demonstrate that, in the plasma membrane, LTPG is relatively
immobile and exhibits a complicated recovery, the latter appears linked to the flux of cuticular lipids through the plasma membrane. LTPG accumulates over the long cell walls of stem
epidermal cells and this protein moves when observed over 1 min intervals. I created a GPIlinked
LTPA and demonstrated that it can rescue the ltpg-1 mutation.
I demonstrate that LTPG is required for wax export by associating with the plant cell wall. This is the first experimental evidence linking the lipid transfer function of a plant LTP to a biological role, which in this case is lipid movement through the cell wall to the cuticle.
6
Gatta, A. "Characterisation of a newly identified family of lipid transfer proteins at membrane contact sites." Thesis, University College London (University of London), 2016. http://discovery.ucl.ac.uk/1517331/.
Full textAbstract:
Non-vesicular intracellular lipid traffic is mediated by lipid transfer proteins (LTPs), which contain domains with an internal cavity that can solubilise and transfer lipids. One of the most widespread LTP folds is the Steroidogenic Acute Regulatory Transfer (StART) domain, which forms a hydrophobic pocket, and appears in proteins with different localisations and lipid specificities. The aim of this study was to characterise a new StART-like domain family, which we identified by a bioinformatics approach. I studied aspects of the localisations, functions and structural properties of six StART-like proteins in S. cerevisiae. The yeast StART-like proteins were endoplasmic reticulum (ER)-integral membrane proteins with transmembrane domains, and they localised at membrane contact sites: Lam1p/Lam3p, and Lam2p/Lam4p at junctions between ER and plasma membrane (PM); Lam5p/Lam6p at junctions between the ER and the vacuolar membrane, at nucleus-vacuole junction (NVJ) and at ER-mitochondria contacts. To study their functions, I purified the second StART-like domain of Lam4p, and I identified sterol as its lipid ligand from in vitro binding assays and in a spectroscopy approach with fluorescent ergosterol. We named the whole family LAM for Lipid transfer proteins Anchored at Membrane contact sites. The sterol binding property of the domains was related to a phenotype shared by LAM1, LAM2 and LAM3 delete strains, which showed an increased sensitivity to the sterol-sequestering polyene antifungal drug Amphotericin B (AmB). The two most sensitive strains (lam1∆ and lam3∆), displayed low sphingolipid levels, which is as yet unexplained. All AmB phenotypes were rescued by StART-like domains from the human LAMa, Lam2/4p and Lam5/6p, suggesting that these domains bind sterol. Simultaneous deletion of LAM1, LAM2, and LAM3 significantly reduced the extent of cortical ER-PM contacts, implying that they create the structure of the particularly punctate contact site they target. Finally, I started structural analysis of Lam4S2 to study the mechanism of sterol binding and to confirm our structural model.
7
Goehring, Natalia. "Phosphatidylinositol transfer proteins : does the topology and the stored curvature elastic stress of lipid bilayers regulate membrane-association and lipid abstraction?" Thesis, Imperial College London, 2012. http://hdl.handle.net/10044/1/9277.
Full textAbstract:
Lipids in a bilayer determine the stresses and the topology of the membrane they form. An understanding of the link between lipid composition and biomechanical parameters such as the spontaneous curvature and bending rigidity is key to elucidate the mechanisms behind protein‐membrane interactions. Despite their relatively low abundance in‐vivo, one of the most important class of lipids involved in signalling cascades in cells are the phosphatidylinositols (PIs) lipids. Experimental studies of the effect of phosphatidylinositol lipids upon model and cellular membrane systems remain in their infancy and have not matched the pace of discovery with respect to their role in regulating key cellular processes. Synthesised at the endoplasmic reticulum, one route for the distribution of the PIs to other organelle membranes is via translocation by the phosphatidylinositol transfer proteins (PITP). In order to study the link between membrane composition and PITP function, these proteins have been assayed for their interaction and binding with model membranes containing differing amounts of PI. Corresponding studies of the phase behaviour of these systems have been conducted using Small Angle X‐ray scattering specifically investigating the influence PI has in a bilayer membrane formed by dioleoylphosphatidylcholine, as well as in an inverted hexagonal phase formed by dioleoyl‐phosphatidylethanolamine. Additionally, a novel platform based upon a BODIPY fluorescent probe is presented, which is able to sense the stored stresses within lipid bilayers, and whose measurements are correlated these with the make‐up of membranes.
8
Wright, Jenny R. "Investigation of protein-induced formation of lipid domains and their dynamics using fluorescence energy transfer /." Electronic version (PDF), 2005. http://dl.uncw.edu/etd/2005/wrightj/jennywright.pdf.
Full text
9
Ahlsén, Hanna. "The Effects of Abiotic Stress on Alternative Splicing in Non-specific Lipid Transfer Proteins in Marchantia polymorpha." Thesis, Linköpings universitet, Biologi, 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-148937.
Full textAbstract:
Due to global warming, our planet will experience more extreme weather conditions. Plants can protect themselves against these abiotic stress conditions with their stress response, which includes alternative splicing of certain genes. Alternative splicing is a post-transcriptional process where a single gene gives rise to different mRNAs, which in turn produces different proteins. In plants, this is usually done by intron retention. One type of protein that may be involved in this stress response are the non-specific lipid transfer proteins (LTPs). Indeed, evidence of intron retention has been found in the LTP genes in the liverwort Marchantia polymorpha, called MpLTPd. To investigate whether this alternative splicing is caused by abiotic stress or not, I subjected the moss to two different types of stress trials, drought and cold, and compared the general expression of the intron in MpLTPd2 and MpLTPd3 from the stressed samples to samples from a moss grown under normal conditions. I found that the expression of the intron did change in the stressed moss, but none of the differences were significant. This suggests that alterative splicing in MpLTPd2 and MpLTPd3 is not caused by cold and drought and that the intron-containing protein plays no role in the protection of M. polymorpha against abiotic stress.
10
Jayachandra, Pandiyan Muneeswaran. "A bioinformatics approach to investigate the function of non specific lipid transfer proteins in Arabidopsis thaliana." Thesis, Linköpings universitet, Molekylär genetik, 2010. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-57337.
Full textAbstract:
Plant non specific lipid transfer proteins (nsLTPs) enhance in vitro transfer of phospholipids between membranes. Our analysis exploited the large amount of Arabidopsis transcriptome data in public databases to learn more about the function of nsLTPs. The analysis revealed that some nsLTPs are expressed only in roots, some are seed specific, and others are specific for tissues above ground whereas certain nsLTPs show a more general expression pattern. Only few nsLTPs showed a strong up or downregulation after that the Arabidopsis plant had suffered from biotic or abiotic stresses. However, salt, high osmosis and UV-B radiation caused upregulation of some nsLTP genes. Further, when the coexpression pattern of the A.thaliana nsLTPs were investigated, we found that there were several modules of nsLTP genes that showed strong coexpression indicating an involvement in related biological processes. Our finding reveals that the nsLTPs gene was significantly correlated with lipase and peroxidase activity. Hence we concluded that the nsLTPs may play a role in seed germination, signalling and ligning biosynthesis.
11
Abdullah, Syed Umer. "Structural determinants of stability to proteolysis, processing and impact on allergenic potential of non-specific lipid transfer proteins." Thesis, University of East Anglia, 2012. https://ueaeprints.uea.ac.uk/41974/.
Full textAbstract:
Lipid transfer proteins (LTPs) are a class of low molecular weight hydrophobic conserved proteins comprising four intramolecular disulphide bonds making the structure very resistant to proteolysis and harsh food processing conditions. These proteins are identified as strong allergens sensitizing through the gut and share epitopes with LTPs from closely related species. Peach LTP, Pru p 3 is the primary sensitizer in the Mediterranean area being the most frequent food allergen. Wheat LTP, Tri a 14 is a relatively weak allergen with a very low prevalence. The study here compares the structural properties of these proteins and their resistance to various digestive and processing processes. Ligand binding experiments showed that Pru p 3 binds to ligands more strongly than Tri a 14. The gastroduodenal digestion of these LTPs revealed that both are stable to gastric digestion and while Pru p 3 is susceptible to duodenal digestion, Tri a 14 digestion is negligible. Ligand binding did not affect the digestibility of Pru p 3 but improved the duodenal digestibility of Tri a 14. The IgE binding studies using sera from peach allergic individuals confirmed that both Pru p 3 and its digestion fragments in the presence and absence of ligand were IgE reactive. Model processing conditions were employed to treat these LTPs. It was found that heat treatment destroys the secondary structure of Pru p 3 at 121°C and slightly affects that of Tri a 14. Heat treatment also increased the susceptibility of Pru p 3 to gastric digestion while Tri a 14 was less affected. The IgE binding studies showed that heat treatment of Pru p 3 appeared to reduce its IgE recognition while its digestion fragments lost all of their IgE reactivity. To investigate the effect of the food matrix on the digestibility of these LTPs, peach peel containing Pru p 3 and wheat flour containing Tri a 14 were digested under simulated conditions. It was found that they were resistant to proteolysis in their native matrices. Effect of heat treatment to the food matrix again confirmed that both of these proteins were more stable to heat in the matrix and were less digestible. In conclusion, this study shows that there are factors in food matrices which enhance structural stability of LTPs to both processing and digestion. Thus factors such as the effect of food matrix and effect of processing should be taken into account in assessing the allergenic risk posed by foods and not simply rely on data from purified proteins.
12
Fredén, Linnéa. "The Impact of Abiotic Stress on Alternative Splicing in Lipid Transfer Protein in Marchantia polymorpha." Thesis, Linköpings universitet, Biologi, 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-148738.
Full textAbstract:
All plants have a protection against the surrounding environment called a cuticle coating. When this cuticle coating is constructed it is believed that the family of protein called lipid transfer proteins (LTPs) is involved. The LTPs are small and cysteine rich. In Marchantia polymorpha the groups of LTPs called LTPd and LTPg can be found. 8 and 4 in each group respectively. In the genes of LTPd there is an intron placed downstream of the start codon. Firstly, a sequence database search was performed and LTPd2 and LTPd3 were chosen for further experiments in this study. Secondly, a control that the intron was present in the samples were done by preforming a PCR reaction of cDNA from isolated RNA taken from untreated Marchantia polymorpha. A gel electrophoresis of the product was also performed. Lastly, the amount of alternative splicing in LTPd2 and LTPd3 from Marchantia polymorpha after treated with cold and dehydration were studied using quantitative PCR. For the qPCR MpACT and the exon of respective gene were used as references. The ΔCt values and the expression fold (2ΔΔCt) calculated from the qPCR results showed that most of the transcript with introns preserved were upregulated after subjected to stress. Only the intron in MpLTPd2 and MpLTPd3 with MpACT as reference showed a small downregulation after the cold treatment. The intron in MpLTPd3 with MpLTPd3s exon as reference didn’t show any difference. None of the intron transcript in any of the genes on the other hand showed any significant difference in the alternative splicing. This could be because of small sample groups when the test was performed. In conclusion, there were no significant difference in intron expression between treated and control samples. Therefore, nothing can be said about the change in alternative splicing in MpLTPds after cold and dehydration treatments.
13
Kumari, Khushbu [Verfasser], and Dirk [Gutachter] Becker. "The role of lipid transfer proteins (LTPs) during the fertilization process in Arabidopsis thaliana / Khushbu Kumari ; Gutachter: Dirk Becker." Würzburg : Universität Würzburg, 2021. http://d-nb.info/1226669492/34.
Full text
14
Audam, Timothy Ndagi. "Characterization of SIP470, a Family 1 Lipid Transfer Protein and its Role in Plant Stress Signaling." Digital Commons @ East Tennessee State University, 2016. https://dc.etsu.edu/etd/3103.
Full textAbstract:
SIP470, a putative tobacco lipid transfer protein, was identified in a yeast two-hybrid screen to interact with SABP2. SABP2 is a critical role in SA-mediated signaling in tobacco and other plants. In vitro studies using purified recombinant SIP470 confirmed that it is a lipid binding protein. In an attempt to determine its role in mediating stress responses, Arabidopsis T-DNA insertion knockout lines lacking SIP470 homolog were used for the analysis. These mutant plants were defective in basal resistance against microbial pathogens. Expression of defense gene PR-1 was also delayed in these mutant plants. Interestingly, these mutant plants were not defective in inducing systemic acquired resistance. Besides biotic stress, these mutant plants also showed increased susceptibility to abiotic stresses. To directly study the role of SIP470 in tobacco plants, transgenic tobacco lines, with reduced levels of SIP470 expression, were generated using RNAi and transgenic lines overexpressing SIP470 were also generated.
15
Moser, von Filseck Joachim. "Mécanismes du transport lipidique par les protéines ORP/Osh." Thesis, Nice, 2014. http://www.theses.fr/2014NICE4138/document.
Full textAbstract:
Une distribution lipidique hétérogène est essentielle à l’identité et fonction des organelles, mais l’échange par trafic vésiculaire tend à annuler cette distribution. Il existe donc des mécanismes qui assurent l’homéostasie des lipides. Les protéines Osh (S. cerevisiae) et les OSBP-Related Proteins (ORP, H. sapiens), sont des transporteurs de lipides. Osh4 est capable d’échanger de l’ergostérol contre le phosphatidylinositol-4-phosphate (PI4P), présent sur l’appareil de Golgi. Utilisant des outils fluorescents mesurant avec une précision inégalée le transport de stérol et de PI4P, nous démontrons qu’Osh4 transporte du stérol contre son gradient de concentration en utilisant l’énergie d’un gradient de PI4P. Un couplage au métabolisme du PI4P permettrait à Osh4 d’alimenter le Golgi avec du stérol, ainsi créant le gradient de stérol entre ces organelles. La protéine OSBP participe, via sa capacité à connecter la membrane du RE à celle du trans-Golgi, à la création de jonctions entre ces organelles. Nous avons montré qu’OSBP, par échange stérol/PI4P, utilise le PI4P pour transférer du cholestérol au Golgi, mais également pour autoréguler sa capacité à former les jonctions. Osh6 lie la phosphatidylsérine, nous permettant d’étudier un nouveau mécanisme d’échange. Nous avons résolu la structure cristallographique d’un complexe Osh6/PI4P et avons pu observer l’échange de ces deux ligands par Osh6 entre deux membranes. Cette étude nous permet de suggérer que l’échange de PI4P avec divers lipides, via les protéines Osh/ORP, serait un mécanisme général permettant aux cellules de maintenir le gradient lipidique entre le RE et les membranes tardives de la voie sécrétoireAn uneven lipid distribution is essential for the function of eukaryotic organelles. However, exchange of material by vesicular trafficking has a tendency to perturb this distribution; mechanisms must though exist to ensure lipid homeostasis. Osh proteins (S. cerevisiae) and OSBP-Related Proteins (ORPs, H. sapiens), are lipid transfer proteins (LTPs). Osh4 is capable of exchanging ergosterol for phosphatidylinositol 4-phosphate (PI4P), found on the Golgi. Using novel fluorescent tools to measure with unprecedented precision the transport of sterol and PI4P, we find that Osh4 can transport sterol against its concentration gradient using the energy of a PI4P gradient. Coupled to phosphoinositide metabolism, this allows Osh4 to transport sterol to the trans-Golgi and create the sterol gradients observed between these organelles. OSBP participates in the creation of membrane contact sites (MCSs) via its capacity to connect ER membranes to those of the trans-Golgi. We have shown that it uses PI4P for transporting cholesterol from the ER to the trans-Golgi by sterol/PI4P counterexchange, hence also autoregulating its tethering activity. Finally, the identification of phosphatidylserine as a ligand for Osh6 allowed us to analyze the possible extrapolation of the PI4P counterexchange mechanism. We have solved the crystal structure of Osh6 in complex with PI4P and have been able to follow counterexchange of PI(4)P and PS in vitro. Concluding, our studies allow us to suggest a general mechanism for ORP/Osh-mediated counterexchange of PI4P for other lipids to maintain lipid gradients between the ER and late membranes of the secretory pathway
16
Gallo, Alessandra. "Role of non-vesicular secretion in neuronal development." Thesis, Université de Paris (2019-....), 2019. https://theses.md.univ-paris-diderot.fr/GALLO_Alessandra_va.pdf.
Full textAbstract:
La croissance des neurites au cours du développement neuronal nécessite une expansion de la membrane plasmique (MP) via l’insertion de nouveaux lipides et protéines. Cet événement se produit à la suite de la fusion des vésicules de sécrétion avec la MP. Cependant, plusieurs études ont montré que le transfert non-vésiculaire de lipides au niveau des sites de contact entre le réticulum endoplasmique (RE) et la MP joue aussi un rôle dans la croissance des cellules. Des membres de la famille de synaptotagmines étendues (E-Syts) ont été identifiés comme protéines de transfert des lipides dépendantes du Ca2+ au niveau des jonctions RE-MP.Nous avons découvert qu’un nouveau complexe SNARE aux sites de contact RE-MP, composé par Sec22b et Stx1, est impliqué dans la croissance des neurites bien qu’il soit incapable de favoriser la fusion membranaire. Cependant, la manière dont ce complexe participe à l’extension des neurites reste à élucider. Chez la levure, Sec22 interagit avec les protéines de transfert des lipides de la famille OSH, enrichis aux sites de contact RE-MP.Sur la base de ces observations, notre hypothèse est que le transfert non-vésiculaire de lipides induit par E-Syts au niveau des jonctions RE-MP contenant Sec22b pourrait contribuer à la croissance neuronale. L’objectif de ma thèse était d’explorer cette hypothèse. Nous montrons que Sec22b interagit avec E-Syt2 et Stx1 dans les cellules PC12 et avec E-Syt2, E-Syt3 et Stx3 dans les cellules HeLa. L’interaction Sec22b/E-Syt2 dépend du domaine Longin de Sec22b. La surexpression des E-Syts stabilise l’association Sec22b/Stx1, alors que l’inactivation des E-Syts provoque l’effet inverse. La surexpression de E-Syt2 de type sauvage, mais pas des mutants incapables de transférer les lipides ou non fixés au RE, augmentent la formation de filopodes axonaux et la ramification de neurites dans les neurones en développement. Cet effet est inhibé par une neurotoxine clostridiale clivant Stx1, par l’expression du domaine Sec22b Longin et par un mutant Sec22b ayant une extension entre les domaines SNARE et transmembranaire.En conclusion, mes résultats soutiennent l’idée que les sites de contact Sec22b/Stx1 contribuent à l’expansion de la MP via une interaction avec des protéines de transfert de phospholipides comme E-SytsThe growth of neurites during neuronal development requires a massive increase of surface area via the insertion of new proteins and lipids. This event occurs through the fusion of secretory vesicles with the plasma membrane (PM), the final step of the secretory pathway. Recently, non-vesicular transfer of lipids at contacts between endoplasmic reticulum (ER) and PM was shown to contribute to membrane expansion. Members of the ER-integral membrane protein Extended-Synaptotagmin (E-Syt) family have been identified as Ca2+-dependent lipid transfer proteins at ER-PM contact sites, and shown to transfer glycerophospholipids via their lipid binding domains. The laboratory previously found that a novel ER-PM SNARE complex, composed of the ER-resident Sec22b and the neuronal plasmalemmal Stx1, is involved in neurite growth despite being unable to mediate membrane fusion. However, how this complex participates to neurite extension remained to be elucidated. In yeast, Sec22 interacts with lipid transfer proteins of the OSH family, enriched at the ER- PM contacts, supporting a role for Sec22b-populated ER- PM junctions in non-vesicular lipid transport between these bilayers. Based on these observations, our starting hypothesis was that E-Syts-mediated non-vesicular lipid transfer at Sec22b-populated ER-PM contacts, might contribute to neurite growth. The goal of my PhD was to explore this hypothesis with two specific questions: 1-What are the partners of Sec22b complexes which might be involved in the unconventional mechanisms of membrane expansion? 2-What is the mechanism whereby the non-fusogenic SNARE Sec22b/Stx1 complex acts in neuronal development?Here we show that Sec22b interacts with E-Syt2 and Stx1 in PC12 cells and with E-Syt2, E-Syt3 and Stx3 in HeLa cells. Overexpression of E-Syt2 stabilized Sec22b-Stx3 association, whereas silencing of E-Syt2 had the opposite effect. Overexpression of E-Syt2 full length, but not the mutant forms which are unable to transfer lipids or attach to the ER, increased the formation of filopodia particularly in the growing axon. Finally, this effect was inhibited by a clostridial neurotoxin cleaving Stx1, by the expression of Sec22b Longin domain and a by a Sec22b mutant with extended linker between SNARE and transmembrane domains.In conclusion, these results support the hypothesis that Sec22b/Stx1 junctions may contribute to membrane expansion via an interaction with phospholipid transfer proteins like E-Syts
17
Camlioglu, Errol B. "The effect of lipoprotein structure on interlipoprotein lipid transfers by cholesteryl ester transfer protein." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1998. http://www.collectionscanada.ca/obj/s4/f2/dsk2/ftp01/MQ28405.pdf.
Full text
18
Freitas, Gabriela Peres Moraes de, and Gabriela Peres Moraes de Freitas. "Estresse por baixa temperatura em arroz: aspectos moleculares, bioquímicos e fisiológicos." Universidade Federal de Pelotas, 2017. http://guaiaca.ufpel.edu.br:8080/handle/prefix/3973.
Full textAbstract:
Submitted by Maria Beatriz Vieira (mbeatriz.vieira@gmail.com) on 2018-06-11T17:00:10Z
No. of bitstreams: 2
license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5)
resumo_tese_gabriela_peres_moraes_de_freitas.pdf: 25733 bytes, checksum: db07dd8031202634a6a7a48362f489ff (MD5)Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-06-11T18:28:39Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) resumo_tese_gabriela_peres_moraes_de_freitas.pdf: 25733 bytes, checksum: db07dd8031202634a6a7a48362f489ff (MD5)
Made available in DSpace on 2018-06-11T18:28:39Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) resumo_tese_gabriela_peres_moraes_de_freitas.pdf: 25733 bytes, checksum: db07dd8031202634a6a7a48362f489ff (MD5) Previous issue date: 2017-06-23
Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul - FAPERGS
Devido a sua origem tropical, o arroz (Oryza sativa L.) é muito impactado pelo estresse causado pelas baixas temperaturas e consequentemente apresenta grandes perdas de crescimento e produtividade. É o segundo cereal mais consumido no mundo e o decremento na produção causado por eventos de temperatura extrema, pode ocasionar impactos significativos na economia mundial. A sensibilidade e os sintomas das respostas das plantas ao estresse por baixas temperaturas variam com o estádio de desenvolvimento. Observa-se que no estádio inicial ocorre um retardo e diminuição da germinação das sementes e no vegetativo, clorose foliar, baixa estatura e diminuição do perfilhamento. Já no reprodutivo ocorre o aborto e esterilidade das espiguetas, exerção incompleta da panícula, anormalidades no desenvolvimento das anteras, mancha dos grãos e maturação tardia e incompleta dos mesmos. Este trabalho teve como objetivo avaliar nos estádios vegetativo e reprodutivo, os mecanismos adaptativos de resposta ao estresse por baixas temperaturas, a nível molecular, bioquímico e fisiológico. Foram analisados 22 genótipos de arroz, da coleção “mini-core” do USDA para tolerância ao frio, in vitro, através de parâmetros morfológicos. Destes 22, foram selecionados quatro genótipos (dois tolerantes e dois sensíveis), para avaliação, no estádio vegetativo, de trocas gasosas, análises bioquímicas, expressão gênica e de proteínas. As análises de trocas gasosas e expressão gênica foram repetidas durante o estádio reprodutivo. Os primeiros efeitos do estresse por baixa temperatura foram identificados na fotossíntese em todos os genótipos e em ambos os estádios. O perfil bioquímico e dos genes de proteínas de transferência de lipídeos (LTPs), OsGH3-2, OsSRO1a, OsZFP245, OsTPP1 e as proteínas LRR-RLKs, BHLH, GLYI e LTP1, no estádio vegetativo, demonstraram que, apesar dos impactos da baixa temperatura nos genótipos tolerantes, houve um ajuste rápido para que a homeostase celular fosse mantida, mostrando uma clara diferença na expressão gênica entre os genótipos. Foi visualizado também que a expressão dos genes OsLTP7, OsLTP10, fatores de transcrição e genes induzidos por baixa temperatura foram maiores nos genótipos tolerantes, em ambos os tecidos. O estresse por baixa temperatura afetou severamente os parâmetros de produção, tendo uma perda no rendimento dos grãos de 40 e 90%, nos genótipos tolerantes e sensíveis, respectivamente. Assim, este estudo identificou o genótipo Nipponbare como tolerante a baixas temperaturas em ambos os estádios. Os genes LTP, OsGH3-2, OsSRO1a, OsZFP245 e OsTPP1 e as proteínas LRR-RLKs, bHLH, GLYI e LTP1, são bons candidatos para o “screening” de tolerância a baixas temperaturas, no vegetativo, enquanto que as OsLTP7, OsLTP10, OsNAC9, OsNAC10 e OsNAP podem ser usados, com o mesmo fim, no reprodutivo.
Due to its tropical origin, rice (Oryza sativa) is very impacted by cold stress and consequently presents great losses of growth and yield. It is the second most consumed cereal in the world and the decrease in production caused by extreme temperature events can have significant impacts in the world economy. Sensitivity and symptoms of plant responses to stress due to low temperatures vary with the stage of developmental. It is observed that in the first developmental stage there is a delay and decrease of seed germination and on vegetative stage, foliar chlorosis, short stature and decrease of the profile. It is observed that in the first development stage there is a delay and decrease of the germination of the seeds and in the vegetative stage, leaf chlorosis, short stature and reduction of tillering. At reproductive stage, the abortion and sterility of the spikelets, incomplete panicle excretion, abnormalities in the development of the anthers, grain spots and late and incomplete maturation of the grain. The purposed of this study was to evaluate the adaptive mechanisms of response to stress induced by low temperatures at the molecular, biochemical and physiological levels at the vegetative and reproductive stages. Twenty-two rice genotypes from the USDA mini-core collection for cold tolerance, in vitro, were analyzed through morphological parameters. Of these 22, four genotypes (two tolerant and two sensitive) were selected for vegetative stage evaluation of photosynthetic parameters, biochemical and gene and protein expression. Analysis of photosynthetic parameters and gene expression were repeated during the reproductive stage. The first effects of low temperature stress were identified in photosynthesis in all genotypes and in both stages. Biochemical profile and genes of lipid transfer proteins (LTPs), OsGH3-2, OsSRO1a, OsZFP245, OsTPP1 and the LRR-RLKs, BHLH, GLYI and LTP1 proteins at the vegetative stage have demonstrated that, despite impacts of low temperature in the tolerant genotypes, had a fast adjustment for cellular homeostasis to be maintained, showing a clear difference in gene expression between the genotypes. It was also visualized that the expression of OsLTP7, OsLTP10, transcription factors and low temperature genes were higher in the tolerant genotypes in both tissues. Low temperature stress severely affected production parameters, with a yield loss of 40 and 90% in tolerant and sensitive genotypes, respectively. Therefore, this study identified the genotype Nipponbare as possible tolerant to low temperatures in both stages. LTP genes, OsGH3-2, OsSRO1a, OsZFP245 and OsTPP1, and the porteins LRR-RLKs, bHLH, GLYI and LTP1 are good candidates at the screening for cold tolerance at the vegetative stage, whereas OsLTP7, OsLTP10, OsNAC9, OsNAC10 and OsNAP can be used, for the same purpose, at reproductive stage.
19
Garner, K. L. "An investigation into protein and lipid binding by the phosphatidylinositol transfer protein RdgBβ." Thesis, University College London (University of London), 2012. http://discovery.ucl.ac.uk/1359853/.
Full textAbstract:
The phosphatidylinositol transfer proteins (PITPs) are a family of lipid carrier proteins that bind and transfer phosphatidylinositol (PI) and phosphatidylcholine (PC) between membranes. PITPs are commonly involved in phosphoinositide-requiring processes, including phospholipase C and PI 3-kinase signalling, and membrane trafficking. In this study I focus on the uncharacterised soluble PITP, RdgBβ (PITPNC1). The lipid binding and transfer properties of RdgBβ have scarcely been characterised, and the function of RdgBβ is completely unknown. I uncover that RdgBβ interacts with 14-3-3 through its long, disordered C-terminus. RdgBβ is ubiquitinated and subject to rapid degradation in cells, and binding of 14-3-3 via two phosphorylated residues may serve to protect the protein from protease digestion. Whereas RdgBβ binds 14-3-3 under basal conditions, I deduce that, upon stimulation of cells with phorbol ester, RdgBβ binds the Angiotensin II receptor (AT1R)-associated protein, ATRAP, via its N-terminal PITP domain. Others have shown that ATRAP suppresses Angiotensin II signalling by uncoupling AT1R from G proteins and promoting AT1R internalisation. I find that the RdgBβ-ATRAP interaction is blocked by inhibition of protein kinase C or protein synthesis, and may function to re-localise RdgBβ to the membrane in stimulated cells. Unexpectedly, I find that RdgBβ binds PI and phosphatic acid (PA), rather than PI and PC, and that binding of PA is increased by stimulation of cells with GTPγS. Mass spectrometry is used to analyse the molecular species of PI and PA bound by RdgBβ, and reveals that whereas RdgBβ is non-selective in its binding of PI, it selects short-chain monounsaturated or saturated PA species, likely derived from the hydrolysis of PC by phospholipase D.
20
Walter, Stephanie. "The roles of the carboxyl-terminal regions of phospholipid transfer protein (PLTP) and cholesteryl ester transfer protein (CETP) in lipid transfer." Thesis, University of Ottawa (Canada), 2002. http://hdl.handle.net/10393/6262.
Full textAbstract:
In human plasma, there are two lipid transfer proteins that play major roles in lipoprotein metabolism and reverse cholesterol transport. Cholesterol ester transfer protein (CETP) transfers neutral lipids, and is responsible for all of the plasma cholesteryl ester (CE) and triglyceride (TG) transfer activity. Both CETP and phosopholipid transfer protein (PLTP) can facilitate the exchange of phospholipid (PL) among lipoproteins, but only PLTP mediates net mass PL transfer. Residues at the carboxy termini of CETP and PUP have been shown to be essential for protein function. In order to examine the roles of the C-termini of CETP and PUP in determining lipid specificity, we have expressed and characterized chimeric proteins in which the C-termini of PUP and CETP are exchanged. The chimeras CETP1--460/PLTP 445--476 (CP), CETP1--460/PLTP445--476 -mycHIS (CP-HIS), and PLTP1--444/CETP461--476 (PC) were secreted from COS-7 cells, indicating that the overall fold of the proteins was not disturbed. The proteins CP, CPHIS, and PC each had less than 10% of the CE and TG transfer activities of CETP. The proteins CP and CPHIS each had less than 10% of the PLTP-specific PL transfer activity of PUP. These results suggest that the carboxy-termini of CETP and PLTP act in concert with other parts of the molecule to facilitate lipid transfer. Another goal of this research was to produce and characterize a panel of monoclonal antibodies (mAbs) to human PLTP. Attempts to produce mAbs in mice using protein immunization were unsuccessful, but PLTP-specific antibodies were detected in serum from mice immunized with plasmid DNA encoding the human PLTP sequence.
21
Andrews, Shantaya. "Localization of SIP470, a Plant Lipid Transfer Protein in Nicotiana tabacum." Digital Commons @ East Tennessee State University, 2018. https://dc.etsu.edu/etd/3520.
Full textAbstract:
SABP2-interacting protein 470 (SIP470), a non-specific lipid transfer protein (nsLTP), was discovered in a yeast two-hybrid screening using SABP2 as bait and tobacco leaf proteins as prey. SABP2 is an important enzyme in systemic acquired resistance that converts salicylic acid to methyl salicylate. Localization studies are an important aspect to understanding the biological function of proteins. nsLTPs are generally considered apoplastic proteins and has been localized intracellularly and extracellularly. Transient expression shows highest expression of SIP470-eGFP at 2 days post infiltration into Nicotiana benthamiana. Confocal microscopy showed localization near the periphery of the cell. Subcellular localization using differential centrifugation showed that SIP470 is localized in the mitochondria. Mitochondria membranes are rich in lipids and have shown lipid exchange with the endoplasmic reticulum in mammalian systems. Co-localization of SIP470-eGFP+mCherry did not express complete co-localization in the targeted organelles. Co-localization pattern suggests possible localization in the endoplasmic reticulum.
22
Murray, Cathy Maureen. "Regulation of cholesterol ester transfer protein by dietary lipids /." Internet access available to MUN users only, 2003. http://collections.mun.ca/u?/theses,170168.
Full text
23
Sohal, Awinder K. "Studies of a Brassica napus gene encoding a putative lipid transfer protein." Thesis, University of Glasgow, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.284709.
Full text
24
Martinelli, Ana Elisa Marabini. "Papel dos lípides plasmáticos e fatores pró-inflamatórios na fisiopatologia da insuficiência cardíaca." Universidade de São Paulo, 2017. http://www.teses.usp.br/teses/disponiveis/5/5131/tde-07082017-084252/.
Full textAbstract:
Introdução: A Organização Mundial da Saúde estimou em 2015 que 23 milhões de pessoas em todo o mundo sofrem de insuficiência cardíaca (IC), com taxas de mortalidade equivalentes às do câncer. Níveis mais elevados de HDL-colesterol têm sido associados com maior sobrevivência na IC. É consensual que as várias funções protetoras da HDL devem ser exploradas além da concentração de HDL-colesterol. Transferência de lípides para HDL, mediada por proteínas de transferência CETP e PLTP, é uma etapa importante no transporte reverso de colesterol e metabolismo de HDL.,Desenvolvemos um ensaio in vitro para avaliar as transferências de lípides para a HDL, mostrando que esse fenômeno é alterado em várias condições, como na doença arterial coronária, no diabetes mellitus e pelo estilo de vida sedentário. Recentemente, tem sido descrito que a HDL transporta pequenos RNAs não codificadores de proteína, os chamados microRNAs (miRNAs). Alguns miRNAs foram descritos como reguladores críticos do metabolismo das lipoproteínas. O objetivo deste estudo foi comparar lípides plasmáticos, transferência lipídica para HDL, perfil inflamatório, miRNAs relacionados ao metabolismo de lipoproteínas obtidos de pacientes com IC e de pacientes sem IC (sem-IC). Métodos: Quarenta e oito pacientes com IC foram avaliados, 25 em classe funcional NYHA I e II (IC-I/II) e 23 em NYHA III e IV (IC-III/IV), bem como 50 pacientes sem-IC pareados por gênero e idade. Todos os pacientes com IC apresentavam uma fração de ejeção <=40%. Foram determinadas as concentrações plasmáticas de CETP, LCAT, LDL oxidada (LDLox) e atividade de paraoxonase 1 (PON-1). Transferências de lípides para a HDL foi avaliada a partir da incubação de uma nanopartícula artificial com plasma total. A expressão de miRNAs circulantes envolvidos no metabolismo das lipoproteínas também foi analisada. Resultados: Os níveis de colesterol total, LDL e HDL e triglicérides não diferiram entre os três grupos. A concentração da apolipoproteína A-I foi menor no grupo IC-I/II em comparação ao grupo sem-IC (125±23 versus 142±19; p < 0,05), enquanto que a concentração da apolipoproteína B foi menor em ICIII/ IV comparado ao sem-IC (81±35 versus 114±40; p < 0,001). A transferência de colesterol esterificado (5,44±1,76 versus 6,26±0,85), fosfolípides (19,05±2,5 versus 20,21±1,45) e de triglicérides (6,29±2,05 versus 7,40±1,47) foi menor no grupo IC-III/IV do que no grupo sem-IC (p < 0,05). No entanto, não houve diferença nas transferências entre IC-I/II e sem-IC. A concentração de LDLox foi menor em ambos os grupos com IC comparados ao sem-IC (p < 0,0001). A massa de CETP foi menor em IC-III/IV do que em IC-I/II (2,77±1,3 versus 3,78±1,3; p=0,021). A concentração de LCAT e a atividade de PON-1 não foram diferentes entre os grupos. A análise da expressão dos miRNAs circulantes miR-33a, miR-144, miR-185, miR-125, miR-758, miR-26a, miR- 106b, miR-122 e miR-30c, mostrou-se significantemente aumentada nos indivíduos com IC em comparação aos indivíduos sem-IC, ao passo que o miR- 10b foi o único encontrado diminuído na IC comparado com indivíduos sem-IC (p=0,007). Conclusão: Em pacientes com IC mais severa e sintomática da IC, o processo de transferência de lípides para a HDL está deficiente, bem como alguns dos mecanismos que o regulam, e possivelmente estas alterações influenciem no transporte reverso do colesterol e nas funções protetoras da HDL desses pacientesBackground: World Health Organization estimated that there were twentythree million subjects worldwide suffering from heart failure (HF) in 2015, with mortality rates equivalent to those of cancer. Higher HDL-cholesterol levels have been associated with longer survival in HF. It is now consensual that the various protective functions of HDL should be explored beyond HDLcholesterol. Transfer of lipids to HDL, mediated by transfer proteins CETP and PLTP, is an important step in reverse cholesterol transport and HDL metabolism. Previously, we developed an in vitro assay to test those lipid transfers and showed that transfer of cholesterol to HDL is altered in several conditions, such as coronary artery disease (CAD), diabetes and sedentary lifestyle. Recently, HDL transports small non-coding RNA molecule, called micro RNAs (miRNAs). Some miRNA are critical regulators of lipoprotein metabolism. The aim of this study was compare plasma lipids, lipid transfers to HDL, inflammatory profile, miRNAs related to plasma lipids from patients with HF with those from patients with without HF (non-HF). Methods: Forty-eight HF patients were studied, 25 with functional class NYHA I and II (HF I/II) and 23 with NYHA III and IV (HF III/IV), as well as 50 non-HF patients matched for gender, age and BMI. All HF had ejection fraction <= 40%. CETP, LCAT, oxidized LDL (oxLDL) and paraoxonase 1 (PON-1) activity were determined. Transfers of lipids from a donor artificial nanoparticle to HDL was determined by an in vitro assay in which the emulsion was incubated with whole plasma. Expression of circulating miRNAs involved in cholesterol metabolism was also analyzed. Results: Total, LDL and HDL cholesterol and triglycerides did not differ among the 3 groups. Apolipoprotein A-I was lower in NYHA I/II group compared to non- HF (125±23 versus 142±19; p < 0.05) and apo B was lower in NYHA III/IV group compared to non-HF (81±35 versus 114±40, p < 0.001). The transfer of esterified cholesterol (5.44±1.76 versus 6.26±0.85), phospholipids (19.05±2.5 versus 20.21±1.45) and of triglycerides (6.29±2.05 versus 7.40±1.47) to HDL was lower in HF-III/IV than in non-HF (p < 0.05), but lipid transfers were not different between HF-I/II and non-HF. oxLDL was lower in both HF groups compared to non-HF (p < 0.0001). CETP mass was lower in HF-III/IV than in HF-I/II (2.77±1.3 versus 3.78±1.3; p=0.021). LCAT and PON-1 activity was not different among the groups. Regarding to miRNA, miR-33a, miR-144, miR-185, miR-125, miR- 758, miR-26a, miR-106b, miR-122 e miR-30c were significantly increased in HF compared to non-HF subjects, whereas miR-10b was the only one found to be decreased in HF compared to non-HF subjects (p=0.007). Conclusion: In patients with the more severe and symptomatic HF, the lipid transfer to HDL is deficient, as well as some mechanisms that regulate it, and possibly these changes influence reverse cholesterol transport and the protective functions of HDL in these patients
25
Koutb, Mostafa. "Functional analyses on the nonspecific lipid transfer protein (nsLTP) of apple (Malus domestica)." [S.l.] : [s.n.], 2006. http://deposit.ddb.de/cgi-bin/dokserv?idn=979809177.
Full text
26
Read, Jacqueline. "A mechanism of membrane neutral lipid acquisition by the microsomal triglyceride transfer protein." Thesis, Imperial College London, 2001. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.401631.
Full text
27
Wong, L. H. Y. "Analysis of the novel Lipid transfer protein Anchored at Membrane contact sites (LAM) family." Thesis, University College London (University of London), 2017. http://discovery.ucl.ac.uk/1560219/.
Full textAbstract:
Membrane contact sites are dynamic structures where two organelles come into close proximity to regulate and facilitate the flow of material and information between them. One type of inter-organelle communication is lipid exchange, which is essential for membrane maintenance and in response to environmental and cellular stimuli. We recently discovered a new family of Lipid transfer proteins Anchored at Membrane contact sites (LAMs) that is present in all eukaryotes. LAM proteins are integral Endoplasmic Reticulum (ER) proteins containing at least one domain that is structurally similar to the StARkin domain superfamily, a specialised fold that can bind amphipathic ligands such as lipids. The budding yeast, Saccharomyces cerevisiae, has six such proteins: Lam1p-6p. Lam1p-4p are located at contacts between the ER and the plasma membrane (PM), and Lam1p-3p are implicated in retrograde sterol traffic between the ER and PM. The PM contains a high concentration of sterol where it increases rigidity by altering the packing characteristics of the phospholipids in the bilayer. Sterol is also important in the ER, where its levels are low but it is both synthesised and sensed. However, the mechanism by which sterol traffics between the ER and the PM is unknown. This investigation characterises the phenotype of yeast delete LAM strains on Amphotericin B, a sterol sequestering antifungal agent and shows that the conserved StARkin domain of LAM proteins is responsible for resistance against Amphotericin B. Aspergillus fumigatus, a filamentous fungus, has two LAM proteins and the removal of AfLamA causes a severe growth phenotype. Also, in vitro studies indicate that LAM StARkin domains have a clear sterol transfer activity and a mutation that can diminish the function in vivo and in vitro has been identified. These findings present a new candidate protein family for intracellular sterol trafficking.
28
Andrews, Shantaya Biunca, Timothy Audam, and Dhirendra Dr Kumar. "Characterization of SIP470, A Plant Lipid Transfer Protein, and its Role in Plant Defense Signaling." Digital Commons @ East Tennessee State University, 2018. https://dc.etsu.edu/asrf/2018/schedule/44.
Full textAbstract:
Plants are resilient organisms that are continually evolving and continue to withstand an adverse and dynamic world. SABP2-interacting protein (SIP)-470 is a non-specific lipid transfer protein (nsLTP) that was identified in tobacco. SIP470 was discovered during a yeast two-hybrid screening with SABP2, which is an important methyl esterase enzyme which catalyzes the conversion of immobile MeSA into active salicylic acid (SA) during pathogenic challenge. SA activation and mobility allows for immunity to be carried to other, non-infected parts of the plant. This induced responses is called systemic acquired resistance (SAR) and it is a broad spectrum defense. Like many nsLTPs, SIP470 is small and has a predicted characteristic hydrophobic cavity. nsLTPs are found in higher plants and have repeatedly demonstrated protection in biotic stress including disease resistance, and greater resistance to both bacterial and fungal pathogens in overexpressed transgenic lines. This diverse class is also significantly involved in plant adaptation to environmental changes, namely drought, salinity, and freezing, but also in osmotic stress and wounding. Furthermore, nsLTPs are involved in wax metabolism and seed development. Subcellular localization of nsLTPs varies considerably during in vitro and in recent in vivo studies. SIP470 was originally identified in tobacco plants, and therefore, it is important to study its role directly in tobacco plants. SIP470 and eGFP fusion construct has been generated to study the subcellular localization of SIP470 in tobacco cells. SIP470 localization has shown a discontinuous, punctate arrangement around the membrane periphery which is being further verified by subcellular fractionation. Transgenic tobacco lines that are silenced in SIP470 via RNAi have been generated, and these plants are being screened. Overexpressor transgenic lines of SIP470 have been generated and are under the control of an estradiol-inducible promoter. These transgenic lines will be tested for their response in basal resistance and SAR.
29
Vural-Korkut, Senay. "Analysis of the promoter of the barley gene, blt4.9, which encodes a lipid transfer protein." Thesis, University of Newcastle Upon Tyne, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.313553.
Full text
30
Kamenski, Andrei. "Biochemical and structural characterisation of human phosphatidylinositol transfer protein Nir2 and American hookworm lipid binding protein Na-FAR-1." Thesis, University of Glasgow, 2018. http://theses.gla.ac.uk/8997/.
Full textAbstract:
This study focused on biochemical and structural characterisation of two lipid binding proteins: human phosphatidylinositol transfer protein (PITP) Nir2 and American hookworm fatty acid and retinol binding protein (FAR) Na-FAR-1. Nir2 is a large multi-domain PITP that has recently been implicated in phosphoinositide signalling, where it was demonstrated to regulate phosphatidylinositol-4,5-bisphosphate [PI(4,5)P2] homeostasis. Nir2 acts by reciprocally transporting phosphatidylinositol (PI) and phosphatidic acid (PA) between the plasma membrane (PM) and the endoplasmic reticulum (ER), which allows PI(4,5)P2 to be re-synthesised at the PM. Upon cell stimulation, Nir2 translocates to the ER-PM contact sites and is believed to associate with the PM by binding to PA via its C-terminal LNS2 domain. Due to the proposed role of LNS2-PA binding in Nir2 targeting, a detailed investigation of the binding mechanism is desirable, which could help to reveal more details about Nir2 function in the cell. Expression screening of the difficult-to-express Nir2 LNS2 domain yielded a highly-expressed construct that was employed for characterisation of LNS2-PA binding. The data suggested that Nir2 LNS2 binds PA in a specific manner, interacts with both the polar and apolar regions of PA and might associate with the membrane via both hydrophobic and polar interactions. Although the structure of the LNS2 domain could not be determined, several assays are proposed for the identification of LNS2-PA interaction inhibitors that could be used as tool compounds in the investigation of Nir2 and its homologs. Na-FAR-1 is a small lipid binding protein secreted by the human hookworm Necator americanus that infects hundreds of millions of people globally. Na-FAR-1 is known to bind a range of lipid ligands including fatty acids, retinoids and phospholipids, and was proposed to play a role in parasite-host interactions by facilitating nutrient uptake or sequestering lipid signalling molecules in the host tissues. The structure of Na-FAR-1 has been determined previously, but the molecular details of ligand binding by Na-FAR-1 remained unclear. In this study, the high-resolution structure of Na-FAR-1 in complex with its natural ligand oleic acid was determined, and the ligand binding sites were mapped. Furthermore, phospholipid binding by Na-FAR-1 was investigated, and resonance assignment of Na-FAR-1 in complex with PA was carried out, which can be used to obtain the structure of the complex. In addition, Na-FAR-1’s interaction with lysophosphatidic acid was demonstrated in vitro. As lysophosphatidic acid is a mediator of inflammation, the interaction might have important biological implications if it also occurs in vivo.
31
Chan, King-chung Fred, and 陳敬忠. "Functional characterization of StAR-related lipid transfer domain containing 13 (DLC 2) RhoGAP in the nervous system." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2009. http://hub.hku.hk/bib/B43278449.
Full text
32
Chan, King-chung Fred. "Functional characterization of StAR-related lipid transfer domain containing 13 (DLC 2) RhoGAP in the nervous system." Click to view the E-thesis via HKUTO, 2009. http://sunzi.lib.hku.hk/hkuto/record/B43278449.
Full text
33
Bieber, Michael [Verfasser], and Frank [Gutachter] Waller. "Funktionelle Charakterisierung zweier Lipid Transfer Proteine in der Arabidopsis thaliana Pathogenantwort / Michael Bieber. Gutachter: Frank Waller." Würzburg : Universität Würzburg, 2013. http://d-nb.info/1109749643/34.
Full text
34
Chapagai, Danda P. "Biochemical Characterization of SBIP-470 and its role in SA-mediated Signaling in Plants." Digital Commons @ East Tennessee State University, 2014. https://dc.etsu.edu/etd/2428.
Full textAbstract:
Salicylic acid binding protein 2 (SABP2) is known to play a key role in Salicylic acid mediated defense pathway. SBIP-470 is SABP2 interacting protein that might be putatively involved in transfer of lipids. SBIP-470 was cloned without the signal peptide and expressed in E. coli. In vitro lipid binding assay using recombinant SBIP-470 failed to detect lipid binding. In vitro lipid transfer assay showed recombinant SBIP-470 does not transfer phospholipid. Study has shown that SBIP-470 is highly inducible upon infection with viral as well as bacterial pathogens. Induction of SBIP-470 expression upon the TMV infection most likely depends upon the SABP2 while its expression upon non-host bacterial pathogens is most probably inhibited by the SABP2. A study of Arabidopsis knockout mutants (ltp12 mutant and ltp2 mutant) lacking the SBIP-470 homolog genes showed defects in growth phenotype, and they were found susceptible to bacterial pathogens.
35
Lipp, Nicolas-Frédéric. "Fonctionnement des échangeurs phosphatidylsérine / phosphatidylinositol 4-phosphate à l’interface entre le réticulum endoplasmique et la membrane plasmique." Electronic Thesis or Diss., Université Côte d'Azur, 2020. http://www.theses.fr/2020COAZ6027.
Full textAbstract:
Les protéines de transfert lipidique (LTPs) favorisent la solubilisation des lipides et assurent leur distribution entre des membranes biologiques distinctes. Les protéines Osh6p et Osh7p dans les cellules de levure et les protéines ORP5 et ORP8 dans les cellules humaines sont des LTPs homologues qui partagent une fonction commune : elles transportent la phosphatidylsérine (PS) depuis son lieu de synthèse, le réticulum endoplasmique (RE) vers la membrane plasmique où la PS est abondante et essentielle pour les fonctions cellulaires. Pour exécuter ce transport vectoriel, la PS est échangée contre un second lipide – le phosphatidylinositol 4-phosphate (PI(4)P) – qui est ensuite transporté en sens inverse depuis la membrane plasmique vers le RE. Dans la cellule, le PI(4)P est continuellement produit dans la membrane plasmique et dégradé dans le RE, respectivement par des PI 4 kinases et une PI-phosphatase, ce qui permet d'entretenir un gradient de concentration de PI(4)P constant entre les deux membranes. En exploitant ce gradient, Osh6p/Osh7p et ORP5/8 tirent l'énergie nécessaire à l'accumulation de PS dans la membrane plasmique.Avec Osh6p, le laboratoire a observé in vitro que ce mécanisme d’échange de lipides est très rapide et ceci est vérifié dans la levure. Ceci est sans doute suffisant pour approvisionner la membrane plasmique, dont la surface double en un temps de division cellulaire, avec la quantité appropriée de PS. Durant ma thèse, j’ai examiné deux aspects du mécanisme d’échange PS/PI(4)P. Premièrement, j’ai étudié comment Osh6p/Osh7p assurent le transport rapide de ces lipides à l’interface entre deux membranes, l’une étant légèrement anionique (le RE) et l’autre très anionique (la membrane plasmique). Deuxièmement, j’ai étudié ce qui permet à Osh6p et Osh7p de transporter la PS avec précision uniquement entre ces deux membranes dans la levure. Par des approches de biochimie, de reconstitution in vitro et de spectroscopie de fluorescence en temps réel, j’ai pu mettre en évidence, premièrement, qu’Osh6p, en changeant de conformation lors de la capture de la PS ou du PI(4)P, limitait son temps de rétention sur des membranes anioniques lors d’un processus d’échange, ce qui lui permet de l’effectuer rapidement. Deuxièmement, j’ai contribué à montrer qu’Osh6p effectuait des échanges précis, uniquement à l’interface RE/membrane plasmique, en interagissant avec le facteur d’attachement de membranes Ist2p. L’ensemble de ces études améliore notre compréhension des flux lipidiques dans la cellule et ouvre de nouvelles perspectives concernant la régulation de ces machineries essentielles au fonctionnement de la celluleLipid transfer proteins (LTPs) facilitate the solubilization of lipids and ensure their distribution between distinct biological membranes. Osh6p and Osh7p proteins in yeast cells and the ORP5 and ORP8 proteins in human cells are homologous LTPs that share a common function: they transport phosphatidylserine (PS) from its site of synthesis, the endoplasmic reticulum (ER), to the plasma membrane where PS is abundant and essential for a number of cellular functions. To perform this vectorial transport, these proteins exchange PS for a second lipid – phosphatidylinositol 4-phosphate (PI(4)P)– that is next transported in the opposite direction from the plasma membrane to the ER. In the cell, PI(4)P is continuously made in the plasma membrane and hydrolyzed in the ER, respectively by PI 4 kinases and a PI phosphatase, and this maintains a constant concentration gradient between the two membranes. By exploiting this gradient, Osh6p/Osh7p and ORP5/8 derive the necessary energy to contribute to the accumulation of PS in the plasma membrane. About Osh6p, our lab measured in vitro that this exchange mechanism is very fast, and this is also observed in yeast. This is likely sufficient to supply the plasma membrane whose surface area doubles in a cell division time, with the appropriate amount of PS.During my thesis, I examined three aspects of the PS/PI(4)P exchange mechanism. First, I studied how Osh6p/Osh7p ensure rapid lipid transfer at the interface of two membranes, one that is slightly anionic (i.e., the ER) and the other one that is very anionic (i.e., the plasma membrane). Second, I examined what imparts Osh6p and Osh7p with the ability to accurately transport PS solely between these two membranes in yeast. Finally, I examine whether Osh6p and ORP8 differently transfer PS depending on the nature of its acyl-chains.Using biochemical, in vitro reconstitution assays and real-time fluorescence spectroscopy approaches, I demonstrated, first that Osh6p by undergoing a conformation change when encapsulating PS or PI(4)P limits its retention time on anionic membranes during an exchange process, and is therefore very rapid. Second, I contributed to demonstrating that Osh6p performs accurate exchange, only at the ER/plasma membrane interface, by interacting with the membrane tethering factor Ist2p. All these studies improve our understanding of lipid flows in the cell and open new perspectives concerning the regulation of these essential machineries critical for cell function
36
Silva, Renan Gonçalves da. "Análise in silico do gene lipid transfer protein (LTP) de cana-de-açúcar e funcional em transformantes de (Nicotiana tabacum) /." Jaboticabal, 2017. http://hdl.handle.net/11449/151990.
Full textAbstract:
Orientador: Sonia Marli ZingarettiBanca: janete Apparecida Desiderio
Banca: Juliana da Silva Coppede
Resumo: A grande expansão da cultura de cana-de-açúcar pelo território brasileiro leva à necessidade do desenvolvimento de cultivares melhoradas e adaptadas às diferentes condições de clima a que são submetidas. Os estresses bióticos e abióticos são fatores que afetam a produtividade de uma cultura e entre esses, o estresse hídrico assume grande importância em função do regime de chuvas e do aumento de temperatura iminente nos próximos anos. Analisar a expressão de genes em plantas submetidas a estresses pode contribuir de forma expressiva para elucidar as rotas de defesa das plantas, contribuindo sobremaneira para o melhoramento da cultura e o desenvolvimento de novas variedades. O projeto teve como objetivo obter transformantes de Nicotiana tabacum in vitro com o gene LTP (Lipid Transfer Protein) e avaliar sua funcionalidade em relação ao estresse por deficiência hídrica, em casa de vegetação por hidroponia. Feita a seleção da EST com base em resultados anteriores obtidos pelo grupo de pesquisa, posterior análise em bancos de dados, realizou-se a aquisição do clone no Centro de Estocagem de Genes (BCCCenter) e o re-sequenciamento para comprovar sua identidade. Estudos in silico foram realizados através da utilização de softwares de bioinformática e a análise da função do gene foi realizada a partir da transformação genética de Nicotiana tabacum via Agrobacterium tumefaciens. Seis transformantes de N. tabacum com o inserto de interesse LTP foram obtidos e testados quanto à tolerânci... (Resumo completo, clicar acesso eletrônico abaixo)
The great expansion of sugarcane cultivation across Brazilian territory leads to the need to develop better cultivars and adapted to the different climatic conditions that are submitted. The biotic and abiotic stresses are factors that affect the productivity of a crop and among them, the water stress will assume great importance due to the rainfall regime and the increase of the imminent temperature in the next years. Analyzing the expression of genes in stress - stressed plants can contribute in an expressive way to elucidate as plant defense routes, contribute to the improvement of the culture and the development of new varieties. The objective of this project was to obtain transformers of Nicotiana tabacum in vitro with the LTP (Lipid Transfer Protein) gene and to evaluate its functionality in relation to stress due to water deficiency in a greenhouse by hydroponics. We made EST selection based on previous results obtained by a research group, later analysis in databases, performing a clone acquisition in the Gene Storage Center (BCCCenter) and resequencing to prove its identity. Silicon studies were carried out through the application of bioinformatics software and an analysis of the genetic function was performed from the genetic transformation of Nicotiana tabacum via Agrobacterium tumefaciens. Six N. tabacum transformants with the LTP insert of interest were obtained and tested for tolerance to water deficit by induction of different concentrations of mannitol. Transformer tobacco plants showed better phenotypic performance compared to untransformed plant and good readaptation after stress. The T1 generation these plants will be used in studies for the biological and functional verification of the action of the inserted gene, through the Real-Time qPCR technique.
Mestre
37
Souza, Adson Ávila de. "McLTP1 (Morinda citrifolia LIPID TRANSFER PROTEIN 1): efeito antimicrobiano in vitro e atividade protetora sobre a sepse induzida em camundongos." reponame:Repositório Institucional da UFC, 2016. http://www.repositorio.ufc.br/handle/riufc/21817.
Full textAbstract:
SOUZA, Adson Ávila de. McLTP1 (Morinda citrifolia LIPID TRANSFER PROTEIN 1): efeito antimicrobiano in vitro e atividade protetora sobre a sepse induzida em camundongos. 2016. 117 f. Dissertação (Mestrado em Bioquímica)-Universidade Federal do Ceará, Fortaleza, 2016.Submitted by Anderson Silva Pereira (anderson.pereiraaa@gmail.com) on 2017-01-23T21:39:38Z No. of bitstreams: 1 2016_dis_aasouza.pdf: 2447488 bytes, checksum: 29d7cdd8e4951ca5cb0f7bf3bbe1e1f4 (MD5)
Approved for entry into archive by Jairo Viana (jairo@ufc.br) on 2017-01-26T18:13:44Z (GMT) No. of bitstreams: 1 2016_dis_aasouza.pdf: 2447488 bytes, checksum: 29d7cdd8e4951ca5cb0f7bf3bbe1e1f4 (MD5)
Made available in DSpace on 2017-01-26T18:13:44Z (GMT). No. of bitstreams: 1 2016_dis_aasouza.pdf: 2447488 bytes, checksum: 29d7cdd8e4951ca5cb0f7bf3bbe1e1f4 (MD5) Previous issue date: 2016
Microbial resistance is a major public health problem of this century, that has led to increased concern of severe sepsis cases in intensive care units. Due to the nonspecific therapy and the absence of drugs approved for sepsis treatment, new therapeutic approaches has been demanded. Among these approaches, plant antimicrobial peptides have emerged as promising molecules with a potential therapeutic intervention in human health. Recently, our research group isolated a lipid transfer protein type 1 (McLTP1 - UniProt Accession Number: C0HJH5) from Morinda citrifolia L. seeds (noni). This study aimed to evaluate the antibacterial and antifungal potential of McLTP1 against species of clinical concern and investigate its protective effect in mice subjected to sepsis. McLTP1 was isolated from noni seeds crude extract by protein fractionation with trichloroacetic acid 2.5% and ultrafiltration in 30 kDa membrane. McLTP1 antimicrobial properties were evaluated in vitro against planktonic cells and biofilm of Candida spp., gram-positive and gram-negative bacteria species in 96-well plates. In addition, it was evaluated the effect of McLTP1 on bacterial virulence factors and its modulatory properties on clinical antimicrobial drugs. Sepsis in mice was induced by cecal ligation and puncture model, being evaluated the survival rate, body weight and fresh organ weights, haematological parameters and antipyretic effect in animals treated intraperitoneally and orally with McLTP1 (8 mg/kg). Although McLTP1 was not able to inhibit Candida spp. biofilm formation, McLTP1 antifungal activity was observed through growth inhibition of Candida parapsilosis planktonic cells at 25 μg/mL (52.72%; p < 0.05), and through the potentiation effect of amphotericin B (0.06 μg/mL; 28.6% inhibition C. parapsilosis growth) when in the cotreatment with McLTP1 (50 μg/mL; 88.1 % inhibition). Regarding to the antibacterial activity, McLTP1 did not inhibit gram-negative bacteria planktonic cells growth, exhibiting effect only against gram-positive bacteria of Staphylococcus genus, being observed inhibitions on planktonic cells ranging from 18.7% (0.78 μg/mL, p < 0.05) to 98.8% (800 μg/mL; p <0.05) and against biofilms from 34.5% (12.5 μg/mL; p < 0.05) to 73.6% (400 μg/mL). McLTP1 reduced the activity of bacterial virulence factors such as catalase and coagulase, and acted in synergism with the antibiotic oxacillin. In prophylactic and therapeutic treatments at a dose of 8 mg/kg by intraperitoneal and oral route, McLTP1 increased the survival of animals with sepsis, showing antipyretic effect within the first hours after sepsis induction. Moreover, McLTP1 significantly reduced leukocytosis condition observed to the animals of vehicle group with sepsis. In conclusion, McLTP1 showed promising antimicrobial properties and protective effects on animals with sepsis, a new activity for this class of proteins, configured as a therapeutic potential candidate to this syndrome.
A resistência microbiana é um dos principais problemas de saúde pública deste século, e que tem levado ao aumento da preocupação dos quadros graves de sepse em unidades de tratamento intensivo. Diante da inespecificidade da terapia e inexistência de drogas aprovadas no tratamento da sepse, tem-se demandado novas abordagens terapêuticas. Dentre essas abordagens, peptídeos antimicrobianos vegetais têm se destacado como moléculas promissoras com possibilidade de intervenção terapêutica na saúde humana. Recentemente, nosso grupo de pesquisa isolou uma proteína transferidora de lipídeos do tipo 1 (McLTP1 - UniProt Accession Number: C0HJH5) das sementes de Morinda citrifolia L (noni). Este trabalho objetivou avaliar o potencial antibacteriano e antifúngico de McLTP1 sobre espécies de interesse clínico e investigar seu efeito protetor em camundongos submetidos à sepse. McLTP1 foi isolada através do fracionamento de proteínas do extrato total das sementes de noni com ácido tricloroacético 2,5% e ultrafiltração em membrana de 30 kDa. Propriedades antimicrobianas de McLTP1 foram avaliadas in vitro sobre crescimento planctônico e biofilmes de espécies de Candida spp. e de bactérias gram-positivas e gram-negativas em placas de 96 poços. Em adição, foi avaliado o efeito de McLTP1 sobre fatores de virulência bacterianos, e sua atividade moduladora sobre antimicrobianos da prática clínica. A sepse em camundongos foi induzida através do modelo de ligadura e perfuração do ceco, sendo avaliados a taxa de sobrevida, peso corpóreo e peso fresco relativo dos órgãos, parâmetros hematológicos e efeito antipirético em animais tratados por via intraperitoneal e oral com McLTP1 (8 mg/kg). Embora McLTP1 não tenha sido capaz de inibir a formação de biofilmes de Candida spp., a natureza antifúngica de McLTP1 foi observada por meio da inibição do crescimento planctônico de C. parapsilosis na concentração de 25 µg/mL (52,72%; p<0,05), e através da potencialização significativa do efeito de anfotericina B (0,06 μg/mL; 28,6% de inibição do crescimento de C. parapsilosis) quando no cotratamento com McLTP1 (50 μg/mL; 88,1% de inibição). Quanto à atividade antibacteriana, McLTP1 não inibiu o crescimento planctônico de bactérias gram-negativas, exibindo efeito contra bactérias gram-positivas do gênero Staphylococcus, sendo observadas inibições sobre células planctônicas variando entre 18,7% (0,78 μg/mL; p<0,05) e 98,8% (800 μg/mL; p<0,05), e sobre biofilmes em 34,5%, (12,5 μg/mL; p<0,05) a 73,6% (400 μg/mL). McLTP1 reduziu a atividade dos fatores de virulência bacterianos catalase e coagulase, e atuou em sinergismo com o antibiótico oxacilina. Em tratamentos profiláticos e terapêuticos na dose de 8 mg/kg pelas vias intraperitoneal e oral, McLTP1 aumentou a sobrevida de animais com sepse, apresentando efeito antipirético já nas primeiras horas após a indução da sepse. Além disso, McLTP1 reduziu significativamente o quadro de leucocitose observado nos animais com sepse do grupo controle. Portanto, McLTP1 possui propriedades antimicrobianas e efeito protetor sobre animais com sepse promissores, fato este inédito para essa classe de proteínas, configurando-se como candidato terapêutico potencial desta síndrome.
38
Chen, Pak-lam Sammy. "Influence of microsomal triglyceride transfer protein (MTP) gene polymorphism on plasma lipids and lipoproteins in southern Chinese." Click to view the E-thesis via HKUTO, 2003. http://sunzi.lib.hku.hk/hkuto/record/B31980922.
Full text
39
Chen, Pak-lam Sammy, and 陳栢林. "Influence of microsomal triglyceride transfer protein (MTP) gene polymorphism on plasma lipids and lipoproteins in southern Chinese." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2003. http://hub.hku.hk/bib/B31980922.
Full text
40
Jülke, Sabine [Verfasser], Jutta [Akademischer Betreuer] Ludwig-Müller, and Christiane [Akademischer Betreuer] Gatz. "Lipid-Transfer-Proteine aus Arabidopsis thaliana - physiologische und molekulare Funktionsanalyse / Sabine Jülke. Gutachter: Jutta Ludwig-Müller ; Christiane Gatz. Betreuer: Jutta Ludwig-Müller." Dresden : Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2013. http://d-nb.info/1068443871/34.
Full text
41
Mandal, Mihir Kumar. "MOLECULAR AND BIOCHEMICAL CHARACTERIZATION OF OLEATE- AND GLYCEROL-3-PHOSPHATE-REGULATED SIGNALING IN PLANTS." UKnowledge, 2012. http://uknowledge.uky.edu/plantpath_etds/3.
Full textAbstract:
Oleic acid (18:1), a monounsaturated fatty acid (FA), is synthesized upon desaturation of stearic acid (18:0) and this reaction is catalyzed by the plastidal enzyme stearoyl-acyl carrier protein-desaturase (SACPD). A mutation in the SSI2/FAB2 encoded SACPD lowers 18:1 levels, which correlates with induction of various resistance (R) genes and increased resistance to pathogens. Genetic and molecular studies have identified several suppressors of ssi2 which restore altered defense signaling either by normalizing 18:1 levels or by affecting function(s) of a downstream component. Characterization of one such ssi2 suppressor mutant showed that it is required downstream of low 18:1-mediated constitutive signaling and partially restores altered defense signaling in the ssi2 mutant. Molecular and genetic studies showed that the second site mutation was in the Nitric Oxide Associated (NOA) 1 gene, which is thought to participate in NO biosynthesis. Consistent with this result, ssi2 plants accumulated high levels of NO and showed an altered transcriptional profile of NO-responsive genes. Interestingly, the partial defense phenotypes observed in ssi2 noa1 plants were completely restored by an additional mutation in either of the two nitrate reductases NIA1 or NIA2. This suggested that NOA1 and NIA proteins participated in NO biosynthesis in an additive manner. Biochemical studies showed that 18:1 physically bound NOA1, in turn leading to its degradation in a protease-dependent manner. In concurrence, overexpression of NOA1 did not promote NO-derived defense signaling in wild-type plants unless 18:1 levels were lowered. Subcellular localization showed that NOA1 and the 18:1-synthesizing SSI2 were present in close proximity within the nucleoids of chloroplasts. Indeed, pathogen- or low 18:1- induced accumulation of NO was primarily detected in the chloroplasts and their nucleoids. Together, these data suggested that 18:1 levels regulate NO synthesis and thereby NO-mediated retrograde signaling between the nucleoids and the nucleus. Since cellular pools of glycerol-3-phosphate (G3P) regulate 18:1 levels, I next analyzed the relationship between G3P and 18:1. Interestingly, unlike 18:1, an increased G3P pool was associated with enhanced systemic immunity in Arabidopsis. This was consistent with G3P-mediated transcriptional reprogramming in the distal tissues. To determine mechanism(s) underlying G3P-conferred systemic immunity, I analyzed the interaction between G3P and a lipid transfer protein (LTP), DIR1. In addition, I monitored localization of DIR1 in both Arabidopsis as well as tobacco. Contrary to its predicted apoplastic localization, DIR1 localized to endoplasmic reticulum and plasmodesmata. The symplastic localization of DIR1 was confirmed using several different assays, including co-localization with plasmodesmatal-localizing protein, plasmolysis and protoplast-based assays. Translocation assays showed that G3P increased DIR1 levels and translocated DIR1 to distal tissues. Together, these results showed that G3P and DIR1 are present in the symplast and their coordinated transport into distal tissues is likely essential for systemic immunity.
In conclusion, this work showed that low 18:1-mediated signaling is mediated via NO, synthesis of which is likely initiated in the plastidal nucleoids. In addition, my work shows that G3P functions as an independent signal during systemic signaling by mediating translocation of the lipid transfer protein, DIR1.
42
Nieuwoudt, Melanie. "LTP1 and LOX-1 in barley malt and their role in beer production and quality." Thesis, Stellenbosch : Stellenbosch University, 2014. http://hdl.handle.net/10019.1/86558.
Full textAbstract:
Thesis (PhD)--Stellenbosch University, 2014.ENGLISH ABSTRACT: Selection of raw materials for a consistent and high quality end product has been a challenge for brewers globally. Various different factors may influence quality and although a great number of methods for malt analysis exist today for the prediction of end product quality, some still do not accurately represent malt performance in beer. This research focussed on determining parameters in malts to predict two of the major beer quality determining factors namely, foam- and flavour stability. Specific biochemical markers in barley malt such as lipid transfer protein 1 (LTP1) lipoxygenase-1 (LOX-1), anti-radical/oxidant potential (AROP), free amino nitrogen and intact protein were determined and used in beer quality prediction from malt character. These biochemical quality predictions were then correlated with the end product beer quality as assessed in sensory analysis trials on micro-brewed beers. Being such a multi-faceted factor in beer, LTP1 have already become an attractive field of study. LTP1 is primarily associated with stable beer foam, as a foam protein in its own right, and acting as a lipid scavenger. This protein is also theorised to play a role in the stability of beer flavour by possibly acting as anti-oxidant. Lastly LTP1 is known to have anti-yeast activity, which could negatively impact fermentation. In this study LTP1 and its lipid bound isoform LTP1b were successfully purified in an economical and easy five step protocol. Both isoforms showed temperature stability at temperatures >90°C and prefer more neutral and basic pH environments. Although the reported antioxidant activity was not observed, both purified LTP1 and LTP1b inhibited lipoxygenase-1 (LOX-1) activity, which is responsible for the enzymatic breakdown of linoleic acid to form 2(E)-nonenal. This is a novel finding that links LTP1 also to flavour stability. LTP1 exhibited anti-yeast activity whereas LTP1b lost most if not all the activity. However, since most of the LTP1 is converted to LTP1b and glycosylated isoforms during the brewing process fermentation will not be greatly influenced, while foam and flavour stability could still be promoted by the presence of LTP1b. Flavour deterioration of the final packaged product is partially due to the enzymatic production of 2(E)-nonenal by LOX-1 and the presence of free oxygen radical species, limited anti-radical/oxidant potential (AROP) and LTP1. The development of two 96-well micro-assays based on the ferrous oxidation-xylenol orange (FOX) assay for the determination of LOX-1 and AROP was successfully accomplished and compared well with established assays. The LOXFOX and AROP-FOX assays were specifically developed for the on-site, high throughput comparative determination of LOX-1 and AROP in malt and other brewery samples. The AROP-FOX and LOX-FOX micro-assays and a number of established assays were used to categorise malts in different predicted quality groups, various biochemical markers were measured which included LOX activity, LTP1 content, FAN values, intact protein concentration and AROP. An excellent trend (R2=0.93) was found between FAN/LOX and LTP1/LOX which also correlated with the novel observation that LOX-1 activity is inhibited by LTP1 at various concentrations. These trends could assist brewers in optimal blending for not only high quality end products but also fermentation predictions. To determine whether these biochemical markers selected for screening in barley malt are predictive of shelf life potential of the end product, sensory trials were performed. Three barley malt cultivars were selected for LOX, AROP, LTP1, protein and FAN content and used in micro-brewery trials at 0 and 3 months and evaluated using sensory analysis. Good correlation was found between the biochemical predictors and sensory trial for the best quality malt and beer. These parameters were therefore highly relevant for predicting shelf life potential, although additional research is required to elucidate the effect of LTP1 and LOX-1 on each other during the brewing process, since it seems that high LOX-1 concentrations could be leading to LTP1 decreases. With this study it is proposed that if more detailed protein or FAN characterisation is used together with the screening of LOX-1, LTP1 and AROP, an more accurate shelf life prediction, based on malt analysis, is possible and with the help of these parameters brewers can simply blend malts accordingly.
AFRIKAANSE OPSOMMING: Die keuse van roumateriaal om 'n konstante eindproduk van goeie kwaliteit te lewer, was nog altyd 'n uitdaging vir brouers wêreldwyd aangesien verskeie faktore 'n invloed het op die kwaliteit van die produk. Alhoewel daar tans verskeie metodes vir moutanalise bestaan wat die eindproduk–kwaliteit voorspel, is daar min wat werklik die eindproduk kwaliteit soos voorspel deur moutanalise verteenwoordig. Hierdie navorsing fokus op die bepaling van mout-eienskappe om twee van die belangrikste bierkwaliteitvereistes, naamlik skuim- en geurstabiliteit te voorspel. Spesifieke biochemiese eienskappe in garsmout soos lipiedtransportproteien-1 (LTP1), lipoksigenase-1 (LOX-1), antioksidant-antiradikaal potensiaal (AROP), vry aminostikstof (FAN) is geïdentifiseer en gebruik in voorspelling van bierkwaliteit vanaf moutkarakter. Hierdie biochemiese kwaliteit voorspellings is dan gekorreleer met die eindproduk soos ge-evalueer d.m.v sensoriese analise op mikro-gebroude bier. Omdat LTP1 soveel fasette in bier beïnvloed, het dit reeds 'n aanloklike studiefokus geword. LTP1 word hoofsaaklik geassosieer met stabiele skuimkwaliteit in bier en tree op as 'n lipiedmop (“lipid scavenger”). Die proteien speel teoreties ook 'n rol in die stabiliteit van bier geur deur moontlik as 'n anti-oksidant op te tree. Laastens is LTP1 bekend vir sy antigis aktiwiteit wat moontlik 'n negatiewe uitwerking op fermentasies het. Gedurende hierdie navorsing is LTP1 en sy lipiedbinding isoform LTP1b suksesvol gesuiwer met 'n ekonomies en eenvoudige 5-stap protokol. Beide isoforme het stabiliteit by temperature >90°C en meer neutrale en basiese pH omgewings getoon. Alhoewel die voorheen gerapporteerde anti-oksidant aktiwiteit vir LTP1 nie bevestig kon word nie, is daar wel gevind dat beide LTP1 en LTP1b, LOX-1, wat verantwoordelik is vir die ensimatiese afbraak van linoleensuur na 2(E)-nonenal, se aktiwiteit inhibeer. Dit is 'n unieke bevinding wat LTP1 ook koppel aan geurstabiliteit. LTP1 het antigis aktiwiteit getoon, maar LTP1b het die meeste, indien nie alle antigis-aktiwiteit verloor. Omdat die meeste van die LTP1's omgeskakel word na LTP1b's en geglikosileerde isoforme tydens die brouproses, sal fermentasie nie beduidend beinvloed word nie, maar die skuim- en geurstabiliteit sal steeds bevorder word deur die blote teenwoordigheid van die LTP1b. Geurverval van die finale verpakte produk is gedeeltelik a.g.v die ensimatiese produksie van 2(E)-nonenal deur LOX-1 en die teenwoordigheid van vry suurstofradikaal spesies, beperkte AROP en LTP1. Die ontwikkeling van twee 96-putjie mikroessaïs, gebasseer op die yster oksidasie-xilenol oranje (FOX) essai vir die bepaling van LOX-1 en AROP, was suksesvol en het goed vergelyk met reeds gevestigde essaïs. Die LOX-FOX en AROP-FOX mikroessaïs is spesifiek ontwikkel vir die residente, hoë deurvloei vergelykende bepaling van LOX-1 en AROP in mout en ander brouery-monsters. Die AROP-FOX en LOX-FOX mikroessaïs en 'n paar gevestigde essaïs is gebruik om moute te kategoriseer in die verskillende voorspelde kwaliteitsgroepe. Die biochemiese merkers wat gemeet is het die volgende ingesluit: LOX aktiwiteit, LTP1 inhoud, FAN waardes, proteïen konsentrasie en AROP. 'n Merkwaardige korrelasie (R2=0.93) is gevind tussen FAN/LOX en LTP1/LOX wat ook ooreenstem met die waarneming dat LOX-1 aktiwiteit onderdruk word deur LTP1 by verskeie konsentrasies. Hierdie korrelasies kan brouers help met optimale versnitting van moute vir, nie net die hoogste kwaliteit eindproduk nie, maar ook vir fermentasie voorspellings. Om te bepaal of hierdie geselekteerde biochemiese merkers in mout die potensieële raklewe van die eindproduk verteenwoordig, is sensoriese evaluerings uitgevoer. Drie gars-mout kultivars is geselekteer o.g.v LOX-, AROP-, LTP1-, proteïen- en FAN-inhoud en gebruik in mikro-brouery proewe en op 0 en 3 maande en is ge-evalueer deur sensoriese analise. Goeie korrelasie is gevind tussen die biochemiese voorspellers en sensoriese evaluering vir die beste kwaliteit mout en bier. Hierdie maatstawwe is daarom uiters relevant vir voorspelling van die potensiele rakleeftyd, alhoewel addisionele navorsing nodig is om die effek van LTP1 en LOX-1 op mekaar gedurende die brouproses te bepaal. Dit blyk dat 'n hoë LOX-1 konsentrasies kan lei tot 'n afname in LTP1. Met hierdie studie word dit voorstel dat, as meer gedetaileerde proteien of FAN karakterisering saam met LOX-1, LTP1, en AROP analise uitgevoer word, 'n meer akkurate raklewe voorspelling moontlik is en met behulp van hierdie parameters kan brouers moute dienooreenkomstig versnit.
43
Charvolin, Delphine. "Études structurales des protéines de transfert de lipides du mais et du blé : caractérisation de l'interaction entre protéine et lipide." Grenoble 1, 1997. http://www.theses.fr/1997GRE10008.
Full textAbstract:
Les proteines de transfert de lipide forment une famille de petites proteines, tres abondantes dans les plantes et qui presentent une forte homologie de sequence. Leur fonction est assez mal connue et plusieurs hypotheses ont ete emises: elles peuvent etre des transporteurs de phospholipides membranaires entre les membranes cellulaires, ou bien des transporteurs des monomeres de cutine vers les couches externes des plantes ; elles peuvent jouer un role dans la defense des plantes ou encore participer aux mecanismes de stockage des graisses lors de la maturation des graines. Les structures de la proteine de transfert de lipide du ble en presence de lyso-myristoyl-phosphatidyl-choline et de la proteine de transfert de lipide du mais, en absence de lipide ont ete resolues par diffraction des rayons x, respectivement a 2,6 et 1,9 angstroms de resolution. Les structures obtenues sont proches des autres structures de cette famille, determinees par rmn ou cristallographie: elles sont formees de quatre helices, stabilisees par quatre ponts disulfure. On observe la formation d'une cavite cylindrique allongee au sein de la proteine du mais cristallisee sans lipide: elle mesure environ 20 sur 3 sur 4 angstroms et est formee d'une zone hydrophobe et d'une zone polaire, limitee par une tyrosine et deux arginines. La structure de la proteine du ble cristallisee en presence du lipide met en evidence que la zone hydrophobe de la cavite fixe deux chaines carbonnees de lipide et que la zone polaire fixe vraisemblablement la tete polaire du lipide, les residus tyrosine et arginine jouant probablement un role fondamental dans l'attache de la tete polaire
44
Lutif, Camila Crasto. "Biochemical characterization and evaluation of cytotoxic and allergenic activity of transferring protein isolate lipid Morinda citrifolia L. seeds (Rubiaceae)." Universidade Federal do CearÃ, 2015. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=15362.
Full textAbstract:
CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel SuperiorThis work reports the biochemical characterization, cytotoxic and allergenic effects of a lipid transfer protein isolated from M. citrifolia seeds (McLTP1), with trypsin and alpha-amylase inhibition properties. McLTP1 was purified with a procedure involving trichloroacetic acid precipitation and gel filtration chromatography. This protein showed significant inhibitory activities against trypsin (767,10 Â 8,36 TIU/mgP), chymotrypsin (25,36 Â 0,86 IU/mgP), papain (65,419 Â 0,152 IU/mgP) and alpha-amylase (24,40%). Atomic force microscopy displayed that McLTP1 oligomerized in tetramers showing a central channel. Fluorescence and CD assays revealed that the McLTP1 structure is highly stable, regardless of pH and temperature levels. In vitro, McLTP1 presented a selective cytotoxic effect to human ovarian cancer cells (OVCAR-8; IC50 of 16,6 μg/mL) and demonstrated hemolytic effect against fresh rabbit red blood cels. Similarly to other non-specific lipid transfer protein reported, McLTP1 showed allergenic properties in mice, being considered as a true food allergen since it was able to sensitize the animals via the gastrointestinal tract.
Morinda citrifolia L. à uma espÃcie nativa do Sudeste da Ãsia intensamente investigada em funÃÃo de suas propriedades terapÃuticas reportadas hà mais de 2.000 anos. Recentemente, uma proteÃna transferidora de lipÃdeos denominada McLTP1 (UniProt Accession Number: C0HJH5) foi isolada de sementes de noni pelo nosso grupo de pesquisa. McLTP1 à uma proteÃna termoestÃvel de massa molecular 9,4 kDa, resistente à proteÃlise e dotada de atividades moduladoras da inflamaÃÃo e da dor pela via oral, promissoras e inÃditas para esse grupo de molÃculas. Este trabalho objetivou caracterizar bioquimicamente McLTP1, bem como avaliar o seu potencial alergÃnico em camundongos, como etapas bÃsicas para o seu uso racional e seguro do ponto de vista farmacolÃgico. Em adiÃÃo, as propriedades terapÃuticas de McLTP1 foram tambÃm ampliadas, atravÃs da investigaÃÃo de seu efeito citotÃxico em diferentes linhagens de cÃlulas tumorais. A proteÃna em estudo foi isolada utilizando o protocolo jà estabelecido, envolvendo as etapas de precipitaÃÃo seletiva de proteÃnas do extrato total das sementes de noni com Ãcido tricloroacÃtico 2,5% e cromatografia de exclusÃo molecular. O ensaio de alergenicidade in vivo foi conduzido apÃs prÃvia aprovaÃÃo pelo Comità de Ãtica para Uso de Animais da Universidade Federal do Cearà e utilizou fÃmeas nulÃparas com massa corporal entre 25 e 30 g. McLTP1 apresentou in vitro atividades inibitÃrias de tripsina (767,10  8,36 UIT/mgP), quimotripsina (25,36  0,86 UI/mgP), papaÃna (65,419  0,152 UI/mgP) e alfa-amilase (24,40%). A atividade inibitÃria de tripsina de McLTP1 foi reduzida significativamente em temperaturas superiores a 37 ÂC, apresentando atividade residual de apenas 5,91% quando aquecida a 100 ÂC por 30 min. Essa atividade foi tambÃm influenciada pelo pH, sendo de apenas 30,13% e 39,05% quando a proteÃna foi incubada em tampÃes de pH 3,0 e 12,0. O padrÃo de oligomerizaÃÃo de McLTP1 demonstrou a formaÃÃo de agregados dimÃricos/tetramÃricos delimitando um canal central de diÃmetro de 4,4 nm. As anÃlises espectroscÃpicas mostraram que McLTP1 apresenta espectro de CD similar Ãquele apresentado por outras proteÃnas transferidoras de lipÃdeos e caracterÃstico de proteÃnas ricas em alfa-hÃlice. Espectro de CD de McLTP1 nÃo mostrou alteraÃÃes significativas em diferentes temperaturas e pHs, corroborando com os dados de estabilidade obtidos anteriormente. Diferentemente, em condiÃÃes redutoras (DTT 1 mM) o espectro de CD mostrou alteraÃÃo na estrutura secundÃria da proteÃna e os mÃnimos e mÃximos de elipticidade molar foram tambÃm alterados na presenÃa de micelas iÃnicas de SDS (10 mM). McLTP1 apresentou atividade citotÃxica seletiva contra cÃlulas de cÃncer de ovÃrio (Ovcar-8; CI50: 16,6 μg/mL), nÃo sendo citotÃxica para as cÃlulas tumorais de cÃlon humano (HCT-116), leucemia humano (HL-60) e glioblastoma humano (SF-295) testadas. McLTP1 foi capaz de promover hemÃlise significativa em hemÃcias de coelho a partir da concentraÃÃo de 0,005 mgP/mL. McLTP1 apresentou potencial efeito alergÃnico in silico e em camundongos imunizados pela via oral, induzindo a sÃntese de anticorpos IgG e IgG1. Tal como descrito na literatura para outras LTPs, anticorpos anti-McLTP1 produzidos em coelho foram tambÃm capazes de reconhecer proteÃnas presentes em extratos de Rosaceae, Cucurbitaceae e na polpa do fruto de noni. Os dados obtidos permitiram caracterizar parcialmente a proteÃna em estudo, bem como avaliar o seu potencial imunogÃnico apÃs administraÃÃo oral. Novos testes serÃo conduzidos objetivando avaliar a importÃncia clÃnica dessas respostas, uma vez que testes de toxicidade demonstraram que McLTP1 nÃo foi capaz de promover reaÃÃes adversas em camundongos, mesmo apÃs administraÃÃo da dose de 8 mg/kg por 28 dias.
45
Jamecna, Denisa. "Une région intrinsèquement désordonnée dans OSBP contrôle la géometrie et la dynamique du site de contact membranaire." Thesis, Université Côte d'Azur (ComUE), 2018. http://www.theses.fr/2018AZUR4229/document.
Full textAbstract:
La protéine OSBP est un transporteur de lipides qui régule la distribution cellulaire du cholestérol. OSBP comprend un domaine PH, deux séquences « coiled coil », un motif FFAT (deux phénylalanines dans un environement acide), et un domaine de liaison de lipides (ORD) à son extrémité C-terminale. Le domaine PH interagit avec le PI(4)P et la petite protéine G Arf1-GTP au niveau du Golgi, alors que le motif FFAT interagit avec la protéine VAP-A, résidente du réticulum endoplasmique (RE). En liant simultanément tous ces déterminants, OSBP stabilise des sites de contact membranaire entre RE et Golgi, permettant ainsi un contre-échange cholestérol / PI(4)P par l'ORD. OSBP contient également une longue séquence N-terminale d’environ 80 aa, intrinsèquement désordonnée, composée principalement de glycine, proline et d'alanine. Nous démontrons que la présence de ce N-terminus désordonné augmente le rayon de Stoke de OSBP tronquée du domaine ORD, et limite sa densité d’association sur la membrane portant le PI(4)P. La protéine dépourvue du N terminus favorise l'agrégation symétrique des liposomes PI(4)P (mimant la membrane du Golgi) par les deux domaines PH du dimère OSBP, alors que la présence de la séquence désordonnée empêche cette association symétrique. De même, nous observons que la distribution d’OSBP sur la membrane de vésicules unilamellaires géantes (GUV) varie selon la présence ou l'absence du N-terminus. En présence de la séquence désordonnée, la protéine est répartie de manière homogène sur toute la surface du GUV, alors que la protéine sans N-terminal a tendance à s'accumuler à l'interface entre deux GUV de type Golgi. Cette accumulation locale ralentit fortement la mobilité de la protéine à l’interface. Un effet similaire du N-terminal sur la dynamique des protéines est observé lorsque l’association de membranes de type ER et Golgi est assuré par des protéines monomériques (dépourvue du coiled coil) en présence de Vap-A. Les résultats de nos expériences in vitro ont été confirmés en cellules vivantes, où la séquence intrinsèquement désordonnée contrôle le recrutement d’OSBP sur les membranes Golgiennes, sa mobilité et sa dynamique d’activité au cours des cycles de transfert de lipides. La plupart des protéines de la famille d’OSBP contiennent des séquences N-terminales de faible complexité, suggérant un mécanisme général de régulationOxysterol binding protein (OSBP) is a lipid transfer protein that regulates cholesterol distribution in cell membranes. OSBP consists of a pleckstrin homology (PH) domain, two coiled-coils, a “two phenylalanines in acidic tract” (FFAT) motif and a C-terminal lipid binding OSBP-Related Domain (ORD). The PH domain recognizes PI(4)P and small G protein Arf1-GTP at the Golgi, whereas the FFAT motif interacts with the ER-resident protein VAP-A. By binding all these determinants simultaneously, OSBP creates membrane contact sites between ER and Golgi, allowing the counter-transport of cholesterol and PI(4)P by the ORD. OSBP also contains an intrinsically disordered ~80 aa long N-terminal sequence, composed mostly of glycine, proline and alanine. We demonstrate that the presence of disordered N-terminus increases the Stoke’s radius of OSBP truncated proteins and limits their density and saturation level on PI(4)P-containing membrane. The N-terminus also prevents the two PH domains of OSBP dimer to symmetrically tether two PI(4)P-containing (Golgi-like) liposomes, whereas protein lacking the disordered sequence promotes symmetrical liposome aggregation. Similarly, we observe a difference in OSBP membrane distribution on tethered giant unilamellar vesicles (GUVs), based on the presence/absence of N-terminus. Protein with disordered sequence is homogeneously distributed all over the GUV surface, whereas protein without N-terminus tends to accumulate at the interface between two PI(4)P-containing GUVs. This protein accumulation leads to local overcrowding, which is reflected by slow in-plane diffusion. The effect of N-terminus is also manifested in monomeric OSBPderived proteins that tether ER-like and Golgi-like membranes in the presence of VAP-A. Findings from our in vitro experiments are confirmed in living cells, where N-terminus controls the recruitment of OSBP on Golgi membranes, its motility and the on-and-off dynamics during lipid transfer cycles. Most OSBP-related proteins contain low complexity N-terminal sequences, suggesting a general effect
46
Gable-Guillaume, Christine. "Développement de vecteurs non viraux, de type lipides cationiques, pour la transfection, in vivo, chez la souris swiss : application à la mucoviscidose (doctorat : sciences de la vie et de la sante)." Brest, 1999. http://www.theses.fr/1999BRES3102.
Full text
47
Liu, Kang-Cheng. "Influence of lipid membrane environment on the kinetics of the cytochrome P450 reductase- cytochrome P450 3A4 enzyme system in nanodiscs." Thesis, University of Manchester, 2017. https://www.research.manchester.ac.uk/portal/en/theses/influence-of-lipid-membrane-environment-on-the-kinetics-of-the-cytochrome-p450-reductase-cytochrome-p450-3a4-enzyme-system-in-nanodiscs(b8ee4e84-1230-40cf-9b98-b5d6f457f54c).html.
Full textAbstract:
The cytochrome P450 enzyme system is a multicomponent electron-transfer chain composed of a haem-containing monooxygenase cytochrome P450 (CYP) and one or more redox partners. Eukaryotic CYPs and their redox partner NADPH-dependent cytochrome P450 oxidoreductase (CPR) are involved in many biological processes. Each protein has one N- terminal membrane anchor domain for location within the endoplasmic reticulum (ER). In mammals, CYPs and CPR are especially abundant in liver cells, where they play important roles in the metabolism of steroids, fatty acids, and xenobiotic compounds including numerous drugs of pharmaceutical importance. Incorporation into lipid membranes is an important aspect of CYP and CPR function, influencing their kinetic properties and interactions. In this thesis, soluble nanometer-scale phospholipid bilayer membrane discs, "nanodiscs", were used as a reconstitution system to study the influence of lipid membrane composition on the activities of the abundant human CYP3A4 and human CPR. Both enzymes were expressed and purified from bacteria, and assembled into functionally active membrane-bound complexes in nanodiscs. Nanodisc assembly was assessed by a combination of native and denaturing gel electrophoresis, and a fluorimetric assay was developed to study CYP3A4 reaction kinetics using 7-benzyloxyquinoline as substrate. Kinetic properties were investigated with respect to different lipid membrane compositions: phosphatidyl choline; a synthetic lipid mixture resembling the ER; and natural lipids extracted from liver microsomes. Full activity of the CYP3A4 system, with electron transfer from NADPH via CPR, could only be reconstituted when both CYP3A4 and CPR were membrane-bound within the same nanodiscs. No activity was observed when CPR and CYP3A4 were each incorporated seperately into naodiscs then mixed together, or when soluble forms of CPR were mixed with pre-assembled CYP3A4-nanodiscs. Thus, assembly of the two proteins within the same membrane was shown to be essential for the function of the CPR-CYP3A4 electron transfer system. Comparison of the reaction kinetics in different membrane compositions revealed liver microsomal lipid to have an enhancing effect both on the activity of the assembled CPR-CYP3A4 nanodisc complex, and on the activity of CPR alone incorporated in nanodiscs, when compared either to the synthetic lipid mixture or to phosphatidyl choline alone. Thus, natural lipids appear to possess properties or include components important for the catalytic function of the CYP system, which are absent from synthetic lipid. Input of electrons, measured by NADPH consumption, exceeded product formation rate by the CPR-CYP3A4 complex in nanodiscs, indicating "leakage" in the electron flow, possibly due to uncoupling of the two enzymes. Uncoupling was shown to occur by developing a novel fluorimetric method using the dye MitSOX to detect superoxide production. The significance of this, and to what extent control of coupling could be a natural means of regulation of the CPR-CYP system, remains to be determined. Thus, phospholipid bilayer nanodiscs prove a powerful tool to enable detailed analysis of the reaction kinetics of membrane-reconstituted CPR-CYP systems, and to allow pertinent questions to be addressed concerning the integral significance of the membrane environment.
48
Laquitaine, Laurent. "Etude de la régulation et du rôle physiologique du gène Ub-LTP1 (Ugni blanc Lipid Transfer Protein 1) : analyse fonctionnelle du promoteur et mise en évidence de facteurs inducteurs." Poitiers, 2004. http://www.theses.fr/2004POIT2356.
Full text
49
Moraes, Gabriela Peres. "Expressão diferencial de genes normalizadores e da família LTPs1, em genótipos de arroz sob estresse salino." Universidade Federal de Pelotas, 2013. http://repositorio.ufpel.edu.br/handle/ri/2000.
Full textAbstract:
Made available in DSpace on 2014-08-20T13:59:06Z (GMT). No. of bitstreams: 1
dissertacao_gabriela_peres_moraes.pdf: 2721394 bytes, checksum: 37931b48d7259651d911f5ce19c45303 (MD5)
Previous issue date: 2013-08-16On the production of rice, the salinization of the soil and the water of irrigation, during the establishment phases its closely related to the variation in the levels of production. Among the main damages caused by saline stress at cellular level, are the disturbances in the plasmatic membrane, being its effects manifested by alterations on the permeability, lipid composition, electrical potential and activity of proteins linked to it. The proteins LTPs ("Lipid Transfer Proteins") are related to the transfer and connection of fatty acids and phospholipids between membranes, in addition to being involved in the modification of the lipid composition and their biogenesis. On this note, the objective of this work was to analyze the expression of ten candidate genes to normalizing for studies of genetic expression and verify the differential expression of eleven genes LTPs in rice seedlings. The genotypes BRS Bojuru (tolerant) and BRS Ligeirinho (sensitive) were exposed to 150mM of NaCl in times 0, 24, 48, 72 and 96 hours. The normalizing gene UBQ10 was the most stable for these experimental conditions tested, while the least stable were IF-4a, Tip41-Like and Cyclophilin, being, therefore not in, the least indicated. Among the analyzed LTPs genes, LTP10 presented a high expression value (70 times more) in the 96 hour of stress, when compared to the control in sensitive genotype. For the BRS Bojuru this same gene kept the expression standards while exposed in different times to the stress. The gene LTP14 showed similar patterns of expression between the genotypes studied. In the beginning of the stress, the level of expression practically unaltered, followed by increase and successive decrease after 72 hrs of salt exposure. After the correlation analysis, it was observed that LTP7 and 14 showed a positive correlation, while LTPs 10,26 and 23 showed negative correlation among genotypes. The phylogenetic tree showed a grouping tendency similar to the nucleotides and amino acids sequences analyzed. Based on the results of this study, it was concluded that the gene UBQ10 is the best normalizing for the experimental conditions tested and the genes LTP10, 23 and 26 can be used as a marker for the assisted selection of rice plants for the saline stress for showing contrasting response of expression between genotypes.
Na orizicultura, a salinização do solo e da água de irrigação, durante as fases de estabelecimento e reprodutiva está intimamente relacionada com a variação nos níveis de produção. Dentre os principais danos causados pelo estresse salino em nível celular, estão os distúrbios na membrana plasmática, sendo seus efeitos manifestados por alterações na permeabilidade, na composição lipídica, potencial elétrico e atividade de enzimas e proteínas ligadas a ela. As proteínas LTPs ( Lipid Transfer Proteins ) estão relacionadas com a transferência e ligação de ácidos graxos e fosfolipídios entre as membranas, além de estarem envolvidas na modificação da composição lipídica e biogênese destas. Desta forma, o objetivo deste trabalho foi analisar a expressão de dez genes candidatos a normalizadores para estudos de expressão gênica e verificar a expressão diferencial de 11 genes LTPs em plântulas de arroz. Os genótipos BRS Bojuru (tolerante) e BRS Ligeirinho (sensível) foram expostos a 150 mM de NaCl nos tempos 0, 24, 48, 72 e 96 horas. O gene normalizador UBQ10 foi o mais estável para estas condições experimentais testadas, enquanto que os menos estáveis foram IF-4α, TIP41-LIKE e Cyclophilin, sendo, portanto, os menos indicados. Dentre os genes LTPs avaliados, LTP10 apresentou alto valor de expressão (70 vezes mais) nas 96 horas de estresse, quando comparado ao controle no genótipo sensível. Para BRS Bojuru esse mesmo gene manteve o padrão de expressão durante os tempos de exposição ao estresse. O gene LTP14 apresentou padrão de expressão semelhante entre os genótipos estudados. No início do estresse, o nível de expressão manteve-se praticamente inalterado, seguido de aumento e sucessiva diminuição a partir das 72 h de exposição ao sal. A partir da análise de correlação observou-se que LTP7 e 14 apresentaram correlação positiva, enquanto que LTPs 10, 26 e 23 apresentaram correlação negativa entre os genótipos. As árvores filogenéticas mostraram uma tendência de agrupamento semelhante entre as sequências de nucleotídeos e aminoácidos analisadas. Com base nos resultados obtidos, conclui-se que o gene UBQ10 é o melhor normalizador para as condições experimentais testadas e os genes LTP10, 23 e 26 poderão ser utilizados como possíveis marcadores para a seleção assistida de plantas de arroz para o estresse salino por apresentarem resposta de expressão contrastante entre os genótipos.
50
Guzun-Cojocaru, Tatiana. "Peroxydation des lipides émulsionnés et transfert d'ions fer à l'interface huile / eau stabilisée par des protéines de lait : influence des résidus phosphates et de la stabilité du chélate de fer." Thesis, Dijon, 2010. http://www.theses.fr/2010DIJOS012/document.
Full textAbstract:
Le fer ajouté dans des systèmes complexes tels que les aliments induit divers problèmes comme l'oxydation des lipides ou la précipitation d’autres composés présents dans la matrice. Il s’en suit une diminution de sa biodisponibilité et des défauts de goût. L'utilisation de chélates de fer, comme le bisglycinate de fer ou l’EDTA de fer, constitue une voie intéressante : le fer ainsi protégé n’interagirait pas avec la matrice alimentaire. La stabilité des chélates de fer (bisglycinate de fer et NaFe EDTA), supposés peu réactif pour l’initiation de peroxydation, a été attestée sur des modèles d’émulsions huile dans eau stabilisées par du caséinate de sodium ou de la β lactoglobuline. Dans un second temps, le travail a consisté à vérifier la faible influence de ces complexes sur la nature de l’interface huile/eau (étude de la tension et de la rhéologie interfaciales). La stabilité du bisglycinate de fer en solution aqueuse et en présence de β lactoglobuline a également été évaluée en fonction du pH (pH 2, 3,5 et 6,5) par rayons X. Il a ainsi été montré que la nature des protéines formant l’interface huile/eau et le type de sels de fer jouent un rôle crucial sur la stabilité oxydative des émulsions enrichies en fer. L'affinité des protéines de lait pour les ions fer libres est le premier facteur qui contrôle la peroxydation des matières grasses par l’absence de transfert à l'interface huile/eau. La capacité du complexe bisglycinate à retenir les ions fer et donc à limiter la présence d’ions fer libres (ferriques ou ferreux) apparaît comme le second facteur principal à contrôler. La compétition pour les ions fer entre les groupes fonctionnels des protéines et les contre ions de chélates (glycinate ou EDTA) gouverne ainsi l'état d'oxydation du système dans des conditions acide/base proche de la neutralité. En milieu acide, l'oxydation est principalement gouvernée par l’instabilité du complexe bisglycinate de fer et par conséquent la lente libération fer ionique dans la phase aqueuse. Pour résumer, si le complexe de fer n’est pas déstabilisé et que la protéine à l’interface huile/eau ne présente pas de groupe fonctionnel ayant une forte affinité pour le fer, alors la peroxydation des lipides en émulsion est faible. Dans notre démonstration, si une protéine n’est pas phosphorylée et pour des pH proches de 7, alors l’émulsion ne sera pas peroxydée car le fer ne migre pas à l’interface huile/eauIron incorporated into food systems induces oxidation and precipitation. The consequences are a reduced bioavailability and a modification of other food flavor. The iron chelates such as Fe-bisglycinate and Fe-EDTA represent a possibility to avoid side effects, since the iron is protected. The inertety of Fe bisglycinate and NaFe EDTA for lipid peroxidation has been verified in oil-in-water emulsion models stabilized by sodium caseinate or by β lactoglobulin through the following studies: i/ increase of primary and secondary products of oxidation, ii/ change of the properties of the oil/water interface (tension and interfacial rheology), iii/ the stability of the chelate iron (Fe-bisglycinate) in the aqueous phase with β lactoglobulin at different pH (pH 2, 3.5 and 6.5). The oil/water interface stabilized by proteins with phosphate groups has induced peroxidation, which was not observed with proteins containing no phosphate residue. These results demonstrate also that the type of iron salts plays a crucial role in the oxidative stability of emulsions. The ability of the complex to retain iron ion and to avoid “free” ferrous ion is the first important factor to be controlled. The affinity of milk proteins to bind these free iron ions is the second factor that controls the transfer to oil/water interface. To sum up, the competition for iron ions between functional groups of protein and salt counter ions (glycinate, sulfate or EDTA) governs the oxidation state of the system in neutral conditions. In acidic medium, the oxidation is mainly governed by the stability of the complex and the possibility to free iron in bulk