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Journal articles on the topic "Proteïna AFP"

1

Lin, Min, and Michael L. Cleary. "MLL Translocation Partners AF4, AF5q31, and ENL Associate in a Higher Order Protein Complex with CDK9 and Cyclin T1." Blood 106, no. 11 (2005): 99. http://dx.doi.org/10.1182/blood.v106.11.99.99.

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Abstract The Mixed Lineage Leukemia (MLL) gene is frequently involved in chromosomal translocations that cause acute leukemia. More than 40 different genes have been identified as MLL translocation partners, with the expression of corresponding MLL fusion proteins. The MLL protein has histone methyltransferase activity and is required for embryonic development and hematopoiesis. Several proteins have been demonstrated to associate with MLL in a macromolecular complex, which is believed to have chromatin remodeling function. However, the C-terminal SET domain of MLL, which carries the histone methyltransferase activity, is lost in all MLL fusion proteins, thus making the biochemical functions of the fusion proteins unclear. Moreover, the promiscuity of MLL translocation partners, most of them with no known functions, further complicates an understanding of MLL leukemogenic mechanisms. In this study, we purified a protein complex containing AF4, the most common MLL translocation partner, using a combination of conventional column chromatography and immunoaffinity techniques. The AF4 protein complex contains AF5q31 and ENL, two other MLL translocation partners, as well as CDK9 and Cyclin T1, a heterodimer that regulates transcriptional elongation. Gel filtration confirmed that these five proteins co-fractionate with an estimated overall size of 0.8 MDa. All protein-protein interactions were further confirmed by immunoprecipitation-western blotting from K562 cell nuclear extract. To investigate whether these protein-protein interactions are retained in corresponding MLL fusion proteins, immunoprecipitation-western blotting assays were carried out in human leukemia cell lines harboring MLL chromosomal translocations. We found that MLL-AF4, MLL-AF5q31, MLL-ENL and MLL-AF9 each associate with wild type AF4 complex components, including CDK9 and Cyclin T1. In contrast, MLL-AF6 does not associate with any of the AF4 complex components. We propose that the four nuclear MLL translocation partner proteins (AF4, AF5q31, ENL/AF9), whose translocations are found in over 75% of MLL leukemias, associate in a higher order protein complex with CDK9 and Cyclin T1 and thus function in part to regulate transcriptional elongation. The association of CDK9 and Cyclin T1 with the four MLL fusion proteins suggests a common leukemogenic mechanism that may involve transcriptional elongation, which we are currently investigating. Conversely, MLL-cytosolic fusions, e.g. MLL-AF6, appear to function independently of association with the AF4 protein complex, possibly through a homo-dimerization pathway.
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2

Yokoyama, Akihiko, and Hiroshi Okuda. "The Molecular Mechanism of Transcriptional Activation By MLL-AEP Fusion Proteins." Blood 126, no. 23 (2015): 2435. http://dx.doi.org/10.1182/blood.v126.23.2435.2435.

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Abstract Chromosomal translocations generate a variety of mixed lineage leukemia (MLL) fusion genes, which cause aggressive leukemia. Although >70 different fusion partners have been identified, the majority of the cases are caused by the chimeric genes of MLL and a component of the AEP co-activator complex (hereafter referred to as AEP), which comprises of AF4 family proteins (e.g. AF4, AF5Q31), ENL family proteins (e.g. ENL, AF9), and the P-TEFb elongation factor. MLL-AEP fusion proteins constitutively activate their target genes by recruiting AEP components to their target chromatin, whereas wild-type MLL recruits AEP in a context-dependent manner. In the hematopoietic lineage, MLL fusion proteins aberrantly activate a subset of genes implicated in the hematopoietic stem cell (HSC) program, such as HOXA9 and MEIS1. Constitutive expression of these HSC program genes in hematopoietic progenitors has been shown to induce leukemia in a mouse model. It has been speculated that MLL-AEP activates transcription of those HSC program genes by aberrantly activating transcription elongation. However, it is largely unclear how AEP activates transcription. Using an extensive structure/function analysis, we revealed that a serine-rich domain of the AF4 family proteins, termed pSER, is an essential functional component of MLL-AEP-dependent gene activation and leukemic transformation. Through biochemical purification, we have identified Selectivity Factor 1 (SL1) as a novel factor associated with the pSER domain. SL1 comprises TBP and four TBP-associated factors (TAF1A, TAF1B, TAF1C, TAF1D), and is known as a core component of the pre-initiation complex (PIC) of RNA polymerase I (RNAP1). In the presence of UBF, SL1 forms a PIC on the promoters of ribosomal RNA genes, to drive RNAP1-dependent transcription. However, its role in RNAP2-dependent transcription was unknown. The initiation of RNAP2-dependent transcription in eukaryotes occurs through the loading of TBP to the promoter, via a direct association with the TATA element or through as-yet-unidentified mechanisms. Our results demonstrate that AEP facilitates the initiation of RNAP2-dependent transcription via the loading of TBP onto the TATA element, through SL1 activity. MLL-AEP fusion proteins utilize this TBP-loading function to activate transcription initiation in leukemic transformation. The wild-type AEP complex activates gene expression in the same manner in the physiological conditions. Taken together, our results unveil a novel role of SL1 as a TBP-loading factor in RNAP2-dependent gene activation, and a previously unknown transcription initiation mechanism involving AEP, which is more important than its transcription elongation activities for leukemic transformation. These findings greatly advance our understanding of the molecular mechanism of MLL fusion-dependent leukemic transformation, which was previously interpreted simply as mis-regulated transcription elongation. Disclosures Yokoyama: Dainipon Sumitomo Pharma Co., Ltd.: Research Funding.
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Lin, Shan, Anetta Ptasinska, Salam Adli Assi, et al. "The Transcriptome Heterogeneity of MLL-Fusion ALL Is Driven By Fusion Partners Via Distinct Chromatin Binding." Blood 128, no. 22 (2016): 576. http://dx.doi.org/10.1182/blood.v128.22.576.576.

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Abstract Chromosome rearrangements involving the Mixed Lineage Leukemia (MLL) gene on chromosome 11q23 account for 15-20% of acute lymphoid leukemia (ALL) and confer poor prognosis. Such rearrangements generate the MLL-fusion proteins, in which the N-terminus of the MLL protein mediating chromatin interactions is fused with one of more than 70 different partner proteins. Proteins that are frequently involved in MLL translocations, including AF4, ENL, AF9 and AF10, were identified as components of the super elongation complex (SEC) or DOT1L complex (an H3K79 histone methyltransferase). Based on these observations, a consensus model of MLL-fusion leukemogenesis has been proposed, which suggests that all fusion proteins bind to the targets of wildtype MLL and lead to the aberrant transcriptional elongation and H3K79 methylation via the recruitment of SEC or DOT1L complex and thus the uncontrolled activation of the target genes. Therefore, regardless of the nature of fusion partners, all MLL-fusion proteins work in a similar fashion by dysregulating the same pathways. Our group has successfully established xenograft models of MLL-AF4 and MLL-AF9 B-ALL using human CD34+ hematopoietic stem and progenitor cells transduced with FLAG-tagged MLL-Af4 or MLL-AF9, which faithfully recapitulate the clinical features of the disease. We generated MLL-Af4 and -AF9 ALL using matched units of human CD34+ cells to directly test the consensus model. Interestingly, although having the same genetic background, the immunophenotype of the two ALL are unique, with CD34 expressed only in MLL-Af4 but not -AF9 cells. The transcriptomes of the two ALL were analyzed by RNAseq and dysregulated genes were defined by comparison with the transcriptome of normal pro-B cells (p≤0.05, fold-change≥2). Strikingly, only 40% of MLL-Af4-regulated genes overlap with those of MLL-AF9. This transcriptome heterogeneity is mirrored in clinical samples, where the gene signature generated from our model leukemia can be utilized to accurately classify patient samples in unsupervised hierarchical clustering analysis, with MLL-AF4 patient samples readily distinguishable from MLL-AF9 samples. To identify the mechanisms accounting for this heterogeneity, we performed ChIP-seq analysis using anti-FLAG antibody to compare the chromatin occupancy of MLL-Af4 and MLL-AF9 in our model ALL cells. The MLL-Af4 ChIP-seq signal displayed a clear correlation with those of published MLL-AF4 ChIP-seq datasets from patient-derived cell lines, in the range of 70-90%, highlighting the faithfulness of our model. Surprisingly, MLL-Af4 shows a distinct genome-wide distribution compared to MLL-AF9, with only 20% of MLL-Af4 peaks and 35% of MLL-AF9 peaks overlapping. In contrast to MLL-Af4 which predominantly binds to promoter regions, MLL-AF9 has a relatively greater enrichment at intra- and inter-genic regions. Intriguingly, MLL-AF9 tends to bind at repetitive sequences in introns, suggesting these repetitive sequences may serve as regulatory elements for gene expression. Integration with RNAseq data reveals a significant association between differentially-expressed MLL-Af4 and -AF9 targets and specific chromatin binding of different MLL-fusion proteins. This data demonstrates that chromatin binding is not solely controlled by the MLL portion of the fusion protein and that differential target recognition of different fusion proteins is one molecular mechanism driving gene expression heterogeneity. To test whether distinct co-factor recruitment by MLL-fusions adds another layer regulating gene expression heterogeneity beyond DNA binding, we purified the core complexes of MLL-Af4 and -AF9 from ALL cells by anti-FLAG immunoprecipitation and analyzed by mass spectrometry. These experiments identified both common and fusion-specific interacting proteins. MLL-Af4 showed a higher affinity with SEC component EAF2 but a lower affinity with DOT1L compared to MLL-AF9, suggesting that MLL-fusions have distinct associations with complex components which may achieve differential gene regulation. In summary, our data question the consensus model of MLL-fusion leukemia and emphasize that MLL-fusion ALL is a heterogeneous disease. These findings have important implications for therapy development as each MLL-fusion leukemia could have its own Achilles' heel and customized therapy may need to be introduced for each type of disease. Disclosures No relevant conflicts of interest to declare.
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Bushweller, John H., Benjamin Leach, Ming-jin Chang, et al. "The Role of the Intrinsically Disordered and Multifunctional AF9 C-Terminal Domain in MLL-AF9 Leukemia,." Blood 118, no. 21 (2011): 3464. http://dx.doi.org/10.1182/blood.v118.21.3464.3464.

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Abstract Abstract 3464 Mixed Lineage Leukemia (MLL) fusion proteins disrupt HOX gene regulation through recruitment of transcriptional elongation factors, leading to acute leukemia. AF9 is one of the most common MLL fusion partners and has roles in normal regulation of HOX genes. AF9 has also been shown to interact with multiple transcriptional regulators, including AF4 family proteins which recruit and activate P-TEFb, suggesting a pivotal role in regulation of transcriptional elongation at HOX loci. The mechanism by which AF9 regulates normal transcription and contributes to dysregulated transcription in MLL-AF9 leukemia is poorly understood. We have recently shown that the AF9 ANC1 Homology Domain (AHD), which is retained in MLL fusion proteins, is intrinsically disordered, i.e. it is unstructured on its own, and recruits AF4 family members by coupled folding and binding. In order to further understand this behavior, we have determined the solution structure of the AF9 AHD in complex with AF4 and identified a point mutation which selectively disrupts the AF4-AF9 interaction. Introduction of this mutation into MLL-AF9 results in loss of HOXA9 upregulation and loss of transformation of hematopoietic progenitor cells, establishing the importance of this interaction for MLL-AF9 leukemia. We propose that AF9 functions as a signaling hub which regulates transcriptional elongation by alternative recruitment of transcriptional regulators. Indeed, the AF9 ANC1 homology domain (AHD) has been shown to also bind the H3K79 methyltransferase Dot1L, human polycomb 3 (hPC3), and the BCL6 corepressor, necessitating a carefully orchestrated choreography of binding partners to regulate transcription. Intrinsic disorder may facilitate dynamic exchange between binding partners to mediate rapid changes of HOX gene expression in both normal tissues, as required for specific differentiation programs, and in leukemia. Our results also validate the AF9-AF4 interaction as a potential target for therapeutic development. Disclosures: No relevant conflicts of interest to declare.
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Lee, Young-Sun, Eunjung Ko, Eileen L. Yoon, et al. "Multiplexed Proteomic Approach for Identification of Serum Biomarkers in Hepatocellular Carcinoma Patients with Normal AFP." Journal of Clinical Medicine 9, no. 2 (2020): 323. http://dx.doi.org/10.3390/jcm9020323.

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Alpha fetoprotein (AFP) has been used as a serologic indicator of hepatocellular carcinoma (HCC). We aimed to identify an HCC-specific serum biomarker for diagnosis using a multiplexed proteomic technique in HCC patients with normal AFP levels. A total of 152 patients were included from Guro Hospital, Korea University. Among 267 identified proteins, 28 and 86 proteins showed at least a two-fold elevation or reduction in expression, respectively. Multiple reaction monitoring (MRM) analysis of 41 proteins revealed 10 proteins were differentially expressed in patients with liver cirrhosis and HCC patients with normal AFP. A combination of tripartite motif22 (Trim22), seprase, and bone morphogenetic protein1 had an area under receiver operating characteristic of 0.957 for HCC diagnosis. Real-time PCR and western blot analysis of the paired tumor/non-tumor liver tissue in HCC revealed a reduced expression of Trim22 in the tumor tissue. Also, serum levels of Trim22 were significantly reduced in HCC patients with normal AFP compared to those with liver cirrhosis (p = 0.032). Inhibition of Trim22 increased cellular proliferation in human hepatoma cell lines, whereas overexpression of Trim22 decreased cellular proliferation in hepatoma cell lines. In conclusion, the combination of three serum markers improved the chance of diagnosing HCC. MRM-based quantification of the serum protein in patients with normal AFP provides the potential for early diagnosis of HCC.
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Chen, Lili, Yuqing Sun, Jingya Wang, Hui Jiang, and Andrew G. Muntean. "Differential Regulation of c-Myc/Lin28 Discriminates Subclasses of Rearranged MLL Leukemia." Blood 126, no. 23 (2015): 163. http://dx.doi.org/10.1182/blood.v126.23.163.163.

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Abstract Rearrangements of the 11q23 locus account for ~70% of infant ALL and ~50% of infant AML1 and about 10% of leukemia overall. The prognosis for 11q23 patients is generally poor, however, outcomes vary depending on the fusion partner2. Rearrangements fuse the N-terminus of MLL with one of >70 different partner genes that includes both nuclear and cytoplasmic proteins. Despite the different intracellular localization of these partner proteins, to date, all studied MLL fusion proteins (MLL-FPs) localize to chromatin in the nucleus and drive aberrant transcriptional activation. Recent seminal work by a number of groups has revealed that several of the most common nuclear translocation partner genes (including AF9, ENL, AFF1 (AF4), AFF4 (AF5q31), AF10, AF17 and ELL) assemble into a transcriptional activation complex that includes p-TEFb and/or the histone H3K79 methyltransferase DOT1l. Translocation of MLL with members of this complex results in deregulated transcriptional activation of target genes. Conversely, oligomerization motifs are necessary for transformation following 11q23 translocation with a cytoplasmic partner. However, these mechanisms fail to explain different survival outcomes observed in patients. Further, the transcriptional programs induced in these diverse 11q23 leukemias are currently not well understood. In this study, we examined the genome wide expression profiles in leukemic cells transformed by several MLL-FPs representative of nuclear translocations [t(9;11) (MLL-AF9), t(10;11) (MLL-AF10) and t(11;19) (MLL-ENL)] or cytoplasmic translocations [t(1;11) (MLL-AF1p), t(6;11) (MLL-AF6) and t(11;17) (MLL-Gas7)]. Leukemia cell lines established from mouse bone marrow cells expressing these MLL-FPs proliferated at different rates and mice transplanted with the transformed cells develop leukemia with different latencies remarkably consistent with differences observed in patients harboring different MLL translocations. To elucidate differences in the gene programs induced by different MLL-FPs we performed genome wide expression profiling by RNA-sequencing. These data demonstrated that while the MLL-AF9 and MLL-ENL fusion proteins induce very similar gene programs, the cytoplasmic fusion proteins (MLL-AF6, MLL-AF1p and MLL-GAS7) all possess unique gene signatures. We then performed a pathway analysis comparing nuclear fusion proteins and cytoplasmic fusion proteins and discovered the Myc transcription factor program as one of the top distinguishing features. Myc overexpression significantly increased the growth rate of slow-growing cells that also had low intrinsic Myc, while the growth rate change of more highly proliferative cells was minimal. While all leukemic cell lines were sensitive to the BET inhibitor JQ1 which regulates c-Myc expression, greater sensitivity was observed in those with low c-Myc expression demonstrating the universal importance of this gene program. The Myc target and micro RNA binding protein Lin28B is also differentially expressed between nuclear and cytoplasmic fusions. Negative regulation of miR-150 by Lin28B was observed in all MLL-FP cell lines, which is necessarily downregulated in 11q23 leukemias. We then investigated another Lin28B microRNA target let-7. Interestingly, let-7g expression was significantly increased in MLL-FP transformed cells associated with the longest disease latency. These data demonstrate that differential activation of the c-Myc/Lin28 program accounts for changes in let-7g expression and is associated with MLL-FP disease latency. These data also suggest that patients harboring different 11q23 rearrangements will respond differentially to therapeutic targeting of c-Myc expression dependent on fusion partner. References: 1. Krivtsov AV, Armstrong SA. MLL translocations, histone modifications and leukaemia stem-cell development. Nature reviews Cancer. 2007;7(11):823-833. 2. Balgobind BV, Raimondi SC, Harbott J, et al. Novel prognostic subgroups in childhood 11q23/MLL-rearranged acute myeloid leukemia: results of an international retrospective study. Blood. 2009;114(12):2489-2496. Disclosures No relevant conflicts of interest to declare.
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Lin, Shan, Roger T. Luo, Mahesh Shrestha, Michael J. Thirman, and James C. Mulloy. "The full transforming capacity of MLL-Af4 is interlinked with lymphoid lineage commitment." Blood 130, no. 7 (2017): 903–7. http://dx.doi.org/10.1182/blood-2017-04-777185.

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Abstract Chromosome rearrangements involving the mixed-lineage leukemia gene (MLL) create MLL-fusion proteins, which could drive both acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML). The lineage decision of MLL-fusion leukemia is influenced by the fusion partner and microenvironment. To investigate the interplay of fusion proteins and microenvironment in lineage choice, we transplanted human hematopoietic stem and progenitor cells (HSPCs) expressing MLL-AF9 or MLL-Af4 into immunodeficient NSGS mice, which strongly promote myeloid development. Cells expressing MLL-AF9 efficiently developed AML in NSGS mice. In contrast, MLL-Af4 cells, which were fully oncogenic under lymphoid conditions present in NSG mice, displayed compromised transformation capacity in a myeloid microenvironment. MLL-Af4 activated a self-renewal program in a lineage-dependent manner, showing the leukemogenic activity of MLL-Af4 was interlinked with lymphoid lineage commitment. The C-terminal homology domain (CHD) of Af4 was sufficient to confer this linkage. Although the MLL-CHD fusion protein failed to immortalize HSPCs in myeloid conditions in vitro, it could successfully induce ALL in NSG mice. Our data suggest that defective self-renewal ability and leukemogenesis of MLL-Af4 myeloid cells could contribute to the strong B-cell ALL association of MLL-AF4 leukemia observed in the clinic.
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So, Chi Wai Eric, Piu Wong, Min Lin, and Michael L. Cleary. "Disease Models and Transformation Mechanisms Mediated by MLL-AF4 Family Oncoproteins in Human Leukemia." Blood 104, no. 11 (2004): 467. http://dx.doi.org/10.1182/blood.v104.11.467.467.

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Abstract The Mixed Lineage Leukemia (MLL) gene codes for a histone methyltransferase that is required for hematopoietic development. As a consequence of chromosomal translocations, MLL is fused with over 40 different genes to yield in-frame fusion proteins in acute leukemias. AF4, the most common fusion partner, accounts for 40% of MLL leukemias. The AF4-related proteins, LAF4 and AF5q31, are also fusion partners for MLL in rare cases of leukemia, whereas a fourth family member, FMR2, is a candidate protein for fragile X mental retardation syndrome. MLL fusions with AF4 family proteins manifest as acute biphenotypic or lymphoblastic leukemias that are associated with a poor prognosis, but the lack of appropriate disease models has hampered progress in understanding their underlying molecular mechanisms. Here we report that each of the MLL-AF4 family fusion proteins (the leukemia associated MLL-AF4, MLL-LAF4, and MLL-AF5q31 as well as an artificial MLL-FMR2) is capable of altering the growth and self-renewal properties of primary murine hematopoietic stem/progenitor cells in a serial methylcellulose culture myeloid replating assay. However, significant differences were observed in their respective oncogenic potentials. Cells transduced with MLL-LAF4, MLL-AF5q31 and MLL-FMR2 rapidly expanded during serial replatings, adapted to growth in liquid culture, and induced leukemias in syngeneic recipient mice within 6 months. Conversely, bone marrow cells transduced with MLL-AF4 yielded modest numbers of blast-like colonies in the third round of plating, only transiently expanded in vitro, and quickly underwent terminal differentiation. MLL-LAF4 and MLL-AF5q31 were also capable of enhancing the self-renewal of transduced cells with B lymphoid progenitor phenotypes. A structure/function analysis demonstrated that the previously reported transcriptional transactivation domains of AF4 family proteins were neither necessary nor sufficient for in vitro transformation. Furthermore, the recently reported AF9 interaction motif that is conserved in all AF4 family proteins was also shown to be dispensable for in vitro transformation. These data indicate that transactivation per se, and interaction with AF9 in particular, are not absolutely required for the oncogenic actions of MLL-AF4 family proteins. Conversely, a minimal transformation domain was mapped to the highly conserved carboxy-terminal homology domain shared among all AF4 family proteins, including the Drosophila homolog Lilliputian, and shown to be necessary and sufficient for transformation. Taken together, our studies establish transformation models for MLL-AF4 family fusion proteins and provide critical mechanistic insights into their underlying molecular mechanisms.
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Choudhry, M. A., and I. J. McEwan. "In vitro regulation of reporter gene transcription by the androgen receptor AF1 domain." Biochemical Society Transactions 32, no. 6 (2004): 1103–6. http://dx.doi.org/10.1042/bst0321103.

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The androgen receptor (AR) is a ligand-activated transcription factor that regulates gene expression in response to the steroids testosterone and dihydrotestosterone. AR-dependent gene expression is likely to play an important role in a number of receptor-associated disorders, such as prostate cancer, spinal bulbar muscular atrophy, male type baldness and hirsutism. The AR contains two transactivation domains, termed AF1 (activation function 1) located in the N-terminus and AF2 (activation function 2) in the C-terminal ligand-binding domain. AF2 exhibits weak transcriptional activity, whereas AF1 is a strong regulator of transcription. Transcriptional regulation by AF1 is thought to be modulated by a number of proteins that interact with this region, and by post-translational modifications. Our focus is on the N-terminal-interacting proteins and their regulation of transcription via interaction with the receptor. To better understand the mechanism of AR-AF1 action, we have reconstituted AR activity in HeLa nuclear extracts using a unique dual reporter gene assay. Multiple LexA-binding sites in the promoter allow transcription to be driven by a recombinant AR-AF1–Lex fusion protein. The findings from initial experiments suggest an increase in transcription initiation and elongation rates by AR-AF1–Lex. The role of protein–protein interactions involving co-activators and basal transcription factors and AR-AF1 activity are discussed.
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Kundu, Arpita, Eric Kowarz, Jennifer Reis, and Rolf Marschalek. "Biology of t(6;11) Fusion Proteins and Their Role in MLL-Rearranged Acute Leukemia Lineage Determination." Blood 134, Supplement_1 (2019): 5033. http://dx.doi.org/10.1182/blood-2019-123070.

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Chromosomal translocations are genetic rearrangements where a chromosomal segment is transferred to a non-homologous chromosome which give rise to novel chimeras. Chromosomal rearrangements play a significant role in the development of acute leukemias (acute lymphoblastic leukemia (ALL) or acute myeloid leukemia (AML)). Chromosomal translocation events occurring at 11q23 involving the KMT2A or Mixed-Lineage Leukemia (MLL) gene (n=102) can be diagnosed in about 5-10% of all acute leukemia patients (Marschalek Ann Lab Med 2016), especially prevalent in infant acute leukemias (up to 70% of cases). Different chromosomal translocation partner genes (such as AF4, AF6, AF9orENL and ELL) account for the majority of leukemia cases and have their genomic breakpoints within a major breakpoint cluster region (BCR intron 9-11; Meyer et. al. Leukemia 2018). Some rearrangements are specifically associated with particular disease phenotype e.g. the majority of ALL patients (~ 90%) are mainly caused by the following gene fusions, MLL-AF4, MLL-AF9, MLL-ENL. We are interested in a rare but yet drastic chromosomal translocation t(6;11)(q27;q23) which fuses KMT2A/MLL to Afadin (AFDN/AF6) gene. This chromosomal rearrangement has a very poor prognosis (survival-rate is ~10%) and is predominantly diagnosed in patients with high-risk AML. In this project, we investigate the molecular consequences of two different MLL-AF6 fusions and their corresponding reciprocal AF6-MLL fusions. MLL-AF6 fusions are mainly occurring within MLL intron 9 to 11 and are associated with an AML disease phenotype, while the same fusion occurring within the minor breakpoints region in MLL intron 21 until exon (ex) 24 are mainly diagnosed with T-ALL (T-cell acute lymphoblastic leukemia) disease phenotype. The molecular mechanism that determines the resulting disease phenotype is yet unknown. Therefore, we cloned all of these t(6;11) fusion proteins in order to investigate the functional consequences of the two different breakpoints (MLLex1-9::AF6ex2-30, AF6ex1::MLLex10-37; MLLex1-21::AF6ex2-30, AF6ex1::MLLex22-37). All 4 fusion genes were introduced into our inducible Sleeping Beauty system (Ivics et. al. Mobile DNA 2010; Kowarz et. al. Biotechnol J. 2015) and stably transfected reporter cell lines. Basically, these 4 fusion proteins differ only in the presence or absence of their Plant homeodomain 1-3/Bromodomain (PHD1-3/BD) domain (see Figure 1). The PHD domain regulates the epigenetic and transcriptional regulatory functions of wildtype MLL. Subsequently, we analyzed gene expression differences by the MACE-Seq (Massive Analyses of cDNA Ends). MACE data revealed fundamental differences in gene expression profiles when analyzing the two different sets of t(6;11) fusion genes. The resulting profiles have similarities to either AML or T-ALL and might give a rational explanation for the different lineages in these t(6;11) patients. Altogether, these results notably indicate that our study will provide a novel insight into this type of high-risk leukemia and subsequently will be useful for developing of novel and appropriate therapeutic strategies against acute leukemia. Disclosures No relevant conflicts of interest to declare.
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Dissertations / Theses on the topic "Proteïna AFP"

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Moreno, Gonçalves Ana Beatriz. "Aplicaciones biotecnológicas del gen "afp" (Antifungical Protein) de "Aspergillus giganteus" para la protección de plantas frente a infección por patógenos." Doctoral thesis, Universitat de Barcelona, 2006. http://hdl.handle.net/10803/1000.

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Las plantas están constantemente sometidas a estreses ambientales y los hongos son sus principales patógenos. Actualmente, el control de las enfermedades que causan se realiza utilizando compuestos químicos, generando impacto en el medio ambiente. <br/><br/>Una alternativa es la obtención de plantas transgénicas resistentes. En un principio, la mayoría de los transgenes provenían de las propias plantas (genes involucrados en las respuestas de densa). Actualmente, y dada su reducida efectividad, se están identificando genes de defensa de otros organismos, como bacterias, insectos, animales y hongos. <br/><br/>En este trabajo se ha evaluado la utilidad de la proteína AFP ("antifungical protein"), producida por el hongo del suelo <i>Aspergillus giganteus</i>, para actuar como agente antifúngico frente a fitopatógenos de geranio y arroz. Es una proteína con una estructura compacta y muy básica, que se secreta al espacio extracelular. Estudios anteriores ya habían demostrado su actividad antifúngica frente al hongo <i>Botrytis cinerea</i>, responsable de la enfermedad "podredumbre gris" en muchas plantas, especialmente las ornamentales. Los resultados revelan una fuerte actividad antifúngica, con inhibición tanto del desarrollo de las hifas como de la germinación de las esporas. Cuando se utiliza en combinación con la proteína cecropina A de lepidóptero, se observa un efecto aditivo entre ambas, lo que puede ser útil para desarrollar estrategias de expresión simultánea de ambos genes en plantas transgénicas. Además, la AFP inhibe el crecimiento de <i>B. cinerea</i> tanto <i>in vitro</i> como <i>in vivo</i>, en plantas de geranio.<br/><br/>Por otra parte, el hongo <i>Magnaporthe grises</i> causa la enfermedad piricularosis en arroz. En este trabajo se ha desarrollado una estrategia para expresar el gen <i>afp</i> de manera inducible en plantas transgénicas de arroz con el fin de obtener plantas resistentes y evitar los posibles efectos negativos de una expresión constitutiva, como gasto metabólico y aceptación por los consumidores. Nuestros estudios indicaron que el promotor de un gen PR de maíz, el gen <i>ZmPR4</i>, es funcional e inducible por el hongo <i>M. grisea</i> en plantas de arroz. Este promotor controla la expresión del gen <i>afp</i> en niveles suficientes para conferir resistencia a la infección por este hongo en plantas transgénicas. Además este promotor no es activo en el endospermo de la semilla del arroz (órgano destinado al consumo), con lo que se evita que el producto del transgén se acumule en este tejido. <br/><br/>Dado el potencial del gen <i>afp</i> para aplicación biotecnológica, se hacía necesario determinar su mecanismo de acción, así como sus posibles efectos sobre células animales o vegetales. Para eso, se han realizado diferentes estudios utilizando el hongo <i>M. grisea</i>. Mediante microscopía electrónica de transmisión y confocal, s eha observado que esta proteína es capaz de formar poros en la membrana del hongo, penetrando en la célula y acumulándose en el núcleo. Además, tiene la propiedad de interaccionar con ácidos nucleicos (DNA o RNA). Estros resultados sugieren que su mecanismo de acción se basa en una combinación de dos actividades: formación de poros en la membrana, e interacción con ácidos nucleicos. Se realizaron también ensayos con células vegetales (protoplastos de arroz) y humanas (células HeLa), que han permitido determinar que la proteína AFP no ejerce efecto nocivo significativo sobre ellas. <br/><br/>Los resultados obtenidos permiten concluir que el gen <i>afp</i> es un buen candidato para ser utilizado como transgén en la protección de plantas de geranio y de arroz frente a los hongos <i>Botrytis cinerea</i> y <i>Magnaporthe grisea</i>, respectivamente. El promotor del gen <i>ZmPR4</i> representa una buena opción para dirigir la expresión de genes antifúngicos en plantas transgénicas de arroz.<br>The mold "Aspergillus giganteus" produces the antifungal (AFP) protein. In this work we have analysed the biotechnological applications of this protein against phythopathogenic fungus ("Botrytis cinerea" and "Magnaporthe grisea"). We also studied mechanism of action of AFP against this last fungus. The results are presented in 3 chapters. <br/><br/>1) "Botrytis" blight caused by "Botrytis cinerea" is one of the most widely distributed disease of ornamental plants, especially geranium. Here, the antifungal properties of AFP against various "B. cinerea" isolates were investigated. AFP showed a strong inhibition of mycelial growth and conidial germination by a fungicidal activity. Microscopic observations revealed reduced hyphal elongation and swollen tips. AFP directly protects geranium leaves against "Botrytis". These results are discussed in relation to the potential of the "afp" gene to enhance crop protection against "B. cinerea". <br/><br/>2) Rice blast, caused by "Magnaporthe grisea", is the most important fungal disease of cultivated rice. We have developed a strategy for creating disease resistance to "M. grisea" by pathogen-induced expression of the "afp" gene in transgenic rice plants. Plants expressing the "afp" gene under the control of the maize "ZmPR4" promoter were generated. Transformants showed resistance to "M. grisea". Our results suggest that pathogen-inducible expression of the "afp" gene in rice plants may be a practical way for protection against the blast fungus. <br/><br/>3) The mechanism of action of AFP against "Magnaporthe grisea" was investigated. AFP was able to produce membrane permeabilization on fungal cells but not on rice or HeLa cells. TEM studies revealed cellular degradation and damage of plasma membranesin AFP-treated fungal cells. AFP was found in the nucleus and gel-retardation experiments confirmed that AFP binds nucleic acids. Together, our results suggest that the combination of fungal cell permeabilization, cell-penetrating ability and nucleic acid-binding activity of AFP determines its antifungal activity.
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Baisden, Joseph M. "AFAP-110 is a cSrc activator." Morgantown, W. Va. : [West Virginia University Libraries], 2003. http://etd.wvu.edu/templates/showETD.cfm?recnum=2766.

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Thesis (Ph. D.)--West Virginia University, 2003.<br>Title from document title page. Document formatted into pages; contains v, 149 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references.
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Cherezova, Lidia Nikolayevna. "Determining the effects of phosphorylation on AFAP-110 function." Morgantown, W. Va. : [West Virginia University Libraries], 2002. http://etd.wvu.edu/templates/showETD.cfm?recnum=2492.

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Thesis (M.S.)--West Virginia University, 2002.<br>Title from document title page. Document formatted into pages; contains v, 105 p. : ill. (some col.). Vita. Includes abstract. Includes bibliographical references.
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Dressel, Frank. "Sequenz, Energie, Struktur - Untersuchungen zur Beziehung zwischen Primär- und Tertiärstruktur in globulären und Membran-Proteinen." Doctoral thesis, Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2008. http://nbn-resolving.de/urn:nbn:de:bsz:14-ds-1222781322751-68621.

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Proteine spielen auf der zellulären Ebene eines Organismus eine fundamentale Rolle. Sie sind quasi die „Maschinen“ der Zelle. Ihre Bedeutung wird nicht zuletzt in ihrem Namen deutlich, welcher 1838 erstmals von J. Berzelius verwendet wurde und „das Erste“, „das Wichtigste“ bedeutet. Proteine sind aus Aminosäuren aufgebaute Moleküle. Unter physiologischen Bedingungen besitzen sie eine definierte dreidimensionale Gestalt, welche für ihre biologische Funktion bestimmend ist. Es wird heutzutage davon ausgegangen, dass diese dreidimensionale, stabile Struktur von Proteinen eindeutig durch die Abfolge der einzelnen Aminosäuren, der Sequenz, bestimmt ist. Diese Abfolge ist für jedes Protein in der Desoxyribonukleinsäure (DNS) gespeichert. Es ist allerdings eines der größten ungelösten Probleme der letzten Jahrzehnte, wie die Beziehung zwischen Sequenz und 3D-Struktur tatsächlich aussieht. Die Beantwortung dieser Fragestellung erfordert interdisziplinäre Ansätze aus Biologie, Informatik und Physik. In dieser Arbeit werden mit Hilfe von Methoden der theoretischen (Bio-) Physik einige der damit verbundenen Aspekte untersucht. Das Hauptaugenmerk liegt dabei auf Wechselwirkungen der einzelnen Aminosäuren eines Proteins untereinander, wofür in dieser Arbeit ein entsprechendes Energiemodell entwickelt wurde. Es werden Grundzustände sowie Energielandschaften untersucht und mit experimentellen Daten verglichen. Die Stärke der Wechselwirkung einzelner Aminosäuren erlaubt zusätzlich Aussagen über die Stabilität von Proteinen bezüglich mechanischer Kräfte. Die vorliegende Arbeit unterteilt sich wie folgt: Kapitel 2 dient der Einleitung und stellt Proteine und ihre Funktionen dar. Kapitel 3 stellt die Modellierung der Proteinstrukturen in zwei verschiedenen Modellen vor, welche in dieser Arbeit entwickelt wurden, um 3D-Strukturen von Proteinen zu beschreiben. Anschließend wird in Kapitel 4 ein Algorithmus zum Auffinden des exakten Energieminimums dargestellt. Kapitel 5 beschäftigt sich mit der Frage, wie eine geeignete diskrete Energiefunktion aus experimentellen Daten gewonnen werden kann. In Kapitel 6 werden erste Ergebnisse dieses Modells dargestellt. Der Frage, ob der experimentell bestimmte Zustand dem energetischen Grundzustand eines Proteins entspricht, wird in Kapitel 7 nachgegangen. Die beiden Kapitel 8 und 9 zeigen die Anwendung des Modells an zwei Proteinen, dem Tryptophan cage protein als dem kleinsten, stabilen Protein und Kinesin, einem Motorprotein, für welches 2007 aufschlussreiche Experimente zur mechanischen Stabilität durchgeführt wurden. Kapitel 10 bis 12 widmen sich Membranproteinen. Dabei beschäftigt sich Kapitel 10 mit der Vorhersage von stabilen Bereichen (sog. Entfaltungsbarrieren) unter externer Krafteinwirkung. Zu Beginn wird eine kurze Einleitung zu Membranproteinen gegeben. Im folgenden Kapitel 11 wird die Entfaltung mit Hilfe des Modells und Monte-Carlo-Techniken simuliert. Mit dem an Membranproteine angepassten Wechselwirkungsmodell ist es möglich, den Einfluss von Mutationen auch ohne explizite strukturelle Informationen vorherzusagen. Dieses Thema wird in Kapitel 12 diskutiert. Die Beziehung zwischen Primär- und Tertiärstruktur eines Proteins wird in Kapitel 13 behandelt. Es wird ein Ansatz skizziert, welcher in der Lage ist, Strukturbeziehungen zwischen Proteinen zu detektieren, die mit herkömmlichen Methoden der Bioinformatik nicht gefunden werden können. Die letzten beiden Kapitel schließlich geben eine Zusammenfassung bzw. einen Ausblick auf künftige Entwicklungen und Anwendungen des Modells.
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Rossell, Jacqueline. "Protein immobilisation for AFM." Thesis, University of Nottingham, 2003. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.404144.

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Johansson, Monika. "The role of nucleoside diphosphate kinase in plant mitochondria /." Uppsala : Dept. of Plant Biology and Forest Genetics, Swedish University of Agricultural Sciences, 2006. http://epsilon.slu.se/200674.pdf.

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Yarawsky, Alexander E. "Reversible assembly and amyloidogenesis of the staphylococcal biofilm protein, Aap." University of Cincinnati / OhioLINK, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1560865959517373.

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Cervantes-Laurean, Daniel. "Preparation and Characterization of Model Conjugates for the Study of Proteins Modified by ADP-ribose." Thesis, University of North Texas, 1992. https://digital.library.unt.edu/ark:/67531/metadc935701/.

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Modification of proteins by ADP-ribose has been shown to be a versatile modification with respect to the amino acid side chain. The results described here will allow the study of the biological importance of ADP-ribose glycation and also allow differentiation on crude extracts between enzymatic modifications from protein ADP-ribose glycation that can occur due to the presence of NAD glycohydrolases.
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Xu, Ping. "Sensing and analyzing unfolded protein response during heterologous protein production :." Access to citation, abstract and download form provided by ProQuest Information and Learning Company; downloadable PDF file, 205 p, 2008. http://proquest.umi.com/pqdweb?did=1555621341&sid=2&Fmt=2&clientId=8331&RQT=309&VName=PQD.

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Ndabambi, Nonkululeko. "Recombinant expression of the pRb- and p53-interacting domains from the human RBBP6 protein for in vitro binding studies." Thesis, University of the Western Cape, 2004. http://etd.uwc.ac.za/index.php?module=etd&amp.

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The aim of this thesis was to produce DNA expression constructs and use them to investigate the feasibility of recombinantly expression proteins for future interaction studies between human RBBP6 and p53 and pRb proteins.
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Books on the topic "Proteïna AFP"

1

Althaus, Felix R., and Christoph Richter. ADP-Ribosylation of Proteins. Springer Berlin Heidelberg, 1987. http://dx.doi.org/10.1007/978-3-642-83077-8.

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Althaus, Felix R., Helmuth Hilz, and Sydney Shall, eds. ADP-Ribosylation of Proteins. Springer Berlin Heidelberg, 1985. http://dx.doi.org/10.1007/978-3-642-70589-2.

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1945-, Richter Ch, ed. ADP-ribosylation of proteins: Enzymology and biological significance. Springer-Verlag, 1987.

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cAMP signaling: Methods and protocols. Humana Press, 2015.

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1959-, Gabai Vladimir L., ed. Heat shock proteins and cytoprotection: ATP-deprived mammalian cells. R.G.Landes, 1996.

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Kabakov, Alexander E. Heat shock proteins and cytoprotection: ATP-deprived mammalian cells. Springer, 1997.

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Kim, Sang Geon. AMPK-S6K1 signaling pathway as a target for treating hepatic insulin resistance. Nova Science, 2009.

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Kim, Sang Geon. AMPK-S6K1 signaling pathway as a target for treating hepatic insulin resistance. Nova Science Publishers, 2010.

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Whitfield, James F. Calcium in cell cycles and cancer. 2nd ed. CRC Press, 1995.

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1949-, Althaus F. R., Hilz H. 1924-, Shall S. 1932-, and International Symposium on ADP-Ribosylation Reactions (7th : 1984 : Vitznau, Switzerland), eds. ADP-ribosylation of proteins. Springer-Verlag, 1985.

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Book chapters on the topic "Proteïna AFP"

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Toftager-Larsen, K. "CARBOHYDRATE MICROHETEROGENEITY OF MATERNAL SERUM AFP IN PREGNANCY." In Pregnancy Proteins in Animals, edited by Jann Hau. De Gruyter, 1986. http://dx.doi.org/10.1515/9783110858167-007.

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Westphal, Ulrich. "Androgen-Binding Protein (ABP)." In Steroid-Protein Interactions II. Springer Berlin Heidelberg, 1986. http://dx.doi.org/10.1007/978-3-642-82486-9_9.

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Masignani, V., M. Pizza, and R. Rappuoli. "Common Features of ADP—Ribosyltransferases." In Bacterial Protein Toxins. Springer Berlin Heidelberg, 2000. http://dx.doi.org/10.1007/978-3-662-05971-5_2.

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Popoff, M. R. "Molecular Biology of Actin-ADP-Ribosylating Toxins." In Bacterial Protein Toxins. Springer Berlin Heidelberg, 2000. http://dx.doi.org/10.1007/978-3-662-05971-5_13.

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Lyubchenko, Yuri L. "AFM Visualization of Protein–DNA Interactions." In Single-molecule Studies of Proteins. Springer New York, 2012. http://dx.doi.org/10.1007/978-1-4614-4921-8_4.

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Barbieri, Joseph T., and Drusilla L. Burns. "Bacterial Toxins that Covalently Modify Eukaryotic Proteins by ADP-Ribosylation." In Bacterial Protein Toxins. ASM Press, 2014. http://dx.doi.org/10.1128/9781555817893.ch15.

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Jacobson, Elaine L., Daniel Cervantes-Laurean, and Myron K. Jacobson. "Glycation of proteins by ADP-ribose." In ADP-Ribosylation: Metabolic Effects and Regulatory Functions. Springer US, 1994. http://dx.doi.org/10.1007/978-1-4615-2614-8_27.

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Unsay, Joseph D., and Ana J. García-Sáez. "AFM to Study Pore-Forming Proteins." In Methods in Molecular Biology. Springer New York, 2018. http://dx.doi.org/10.1007/978-1-4939-8894-5_10.

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Patton, W. A., N. Vitale, J. Moss, and M. Vaughan. "Mechanism of Cholera Toxin Action: ADP-Ribosylation Factors as Stimulators of Cholera Toxin-Catalyzed ADP-Ribosylation and Effectors in Intracellular Vesicular Trafficking Events." In Bacterial Protein Toxins. Springer Berlin Heidelberg, 2000. http://dx.doi.org/10.1007/978-3-662-05971-5_7.

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Palazzo, Luca, Dominic I. James, Ian D. Waddell, and Ivan Ahel. "Studying Catabolism of Protein ADP-Ribosylation." In Methods in Molecular Biology. Springer New York, 2017. http://dx.doi.org/10.1007/978-1-4939-6993-7_26.

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Conference papers on the topic "Proteïna AFP"

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Timmons, Sheila, and Jack Hawiger. "REGULATION OF PLATELET RECEPTORS FOR FIBRINOGEN AND VON WILLEBRAND FACTOR BY PROTEIN KINASE." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644674.

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Positive and negative regulation of platelet receptors for adhesive proteins, fibrinogen (F) and von Willebrand Factor (vWF) determines whether binding of these ligands will or will not take place. We have shown previously that ADP stimulates and cyclic AMP inhibits binding of F and vWF to human platelets. Now we show that positive regulation of F and vWF binding to platelets via the glycoprotein 11b/1111a complex is dependent on platelet Protein Kinase C, a calcium- and phospholipid-dependent enzyme. A potent activator of Protein Kinase C, phorbol-12-myristoyl-13-acetate (PMA) induced saturable and specific binding of F and vWF which was inhibited by synthetic peptides, gamma chain .dodecapeptide (gamma 400-411) and RGDS. The phosphorylation of 47kDa protein (P47), a marker of Protein Kinase C activation in platelets, preceded binding of F and vWF induced with PMA as well as with ADP and thrombin. Sphingosine, an inhibitor of Protein Kinase C, blocked binding of F and vWF to platelets stimulated with PMA, ADP, and thrombin. Inhibition of binding was concentration-dependent and it was accompanied by inhibition of platelet aggregation. Thus, stimulation of Protein Kinase C is required for exposure of platelet receptors for adhesive proteins whereas inhibition of Protein Kinase C prevents receptorexposure. Protein Kinase C fulfills the role of an intraplatelet signal transducer, regulating receptors for adhesive proteins, and constitutes a target for pharmacologic modulation of the adhesive interactions of platelets.
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Gear, LR A., D. Freas, and J. D. Carty. "EARLY (< 5 SEC) PHOSPHORYLATIONS OF PLATELET PROTEINS FOLLOWING ACTIVATION BY ADP AND ADRENALIN, SEPARATELY AND IN COMBINATION." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643640.

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Understanding the earliest events (&lt; 1 sec) in signal transduction of platelets is important, since there is evicenee that “shape change,” aggregation and secretion can all begin within this period. We have employed a guenched-flow approach to study these early events and found that thrombin can induce rapid phosphorylation of myosin light-chain kinase (20K) and a 47K protein (Blood, 67, 1738, 1986). To investigate the role of rapid phosphorylations in platelet activation, we have studied the influence of adrenalin and ADP during early (0.3 to 5 sec) stimulation. Aggregation in washed human platelets was assessed by following the loss of single particles and phosphorylation by analysing 32P-labeled proteins after gel electrophoresis. 15 µM adrenalin (without ADP) did not initiate significant aggregation or phosphorylation of myosin light chain (MLC). Phosphorylation of the 47K protein was increased by 20% at 5 sec. 0.5 µM ADP did not induce significant aggregation, but increased phosphorylation of MLC by 130% and the 47 protein by 20%. The combination of 0.5 µM ADP and 15 uM adrenalin induced significant aggregation by 0,3 sec (7.6%), which increased to 25.6% by 5 sec. Interestingly, MLC or 47K protein phosphorylation was not increased above control levels. However, the phosphorylation of four other proteins (77K, 102K, 140K and 185K), which previously had been very rapid (&lt;1 sec) and reversible with 0.5 µM ADP alone, was now maintained, peaking at 3 sec. 10 µM ADP caused small sustained increases in phosphorylation of the same proteins. Adrenalin also caused rapid increases in the phosphorylation of 27K, 213 and 250K proteins. High levels of ADP (10 µM) only increased the 213 and 250K proteins; therefore the 27K protein appears adrenalin specific. Analysis of these early platelet phosphorylations will help understand how they are linked to initiation and maintenance of aggregation. Supported by NIH HL-27014.
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Fujimura, K., T. Fujimoto, M. Takemoto, K. Oda, S. Maehama, and A. Kuramoto. "INTERACTION OF MEMBRANE GLYCOPROTEIN GPIIb AND Ilia WITH CYTOSKELETAL PROTEINS DURING PLATELET ACTIVATION." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643515.

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Experiments were designed and performed to analyse the cytoskeleton assembly and the interaction of glycoprotein (GP)IIb, IIIa and cytoskeletal proteins during platelet activation. A23187 stimulated 125I labeled platelets were solubilised with Triton X-100 solution and centrifuged. The insoluble fraction were analysed by two dimensional electrophoresis and the soluble fraction were fractionated with 5-25% sucrose gradient centrifugation and analysed by SDS PAGE. In Triton X-100 insoluble fraction, high molecular weight protein fraction(MW &gt; 106) was present after stimulation which were consisted of actin binding protein(ABP), myosin heavy chain(MHC), actin and GPIIb and IIIa. And some of the ABP and MHC formed dimer. ABP and actin in this fraction were decreased with 1 mM CaCl2 treatment but the reduction of ABP was inhibited by leupeptm. In Triton X-100 soluble fraction after stimulation, some of the ABP, MHC, P235 protein, actin and small amount of GPIIb, IIIa were sedimented in the same high density fraction but most proteins were sedimented as a monomer form or GPIIb-IIIa complex form. The GPIIb, IIIa incorporation in high molecular weight protein fraction or high density fraction was absent in Ca++ chelating condition or the presence of competitive fibrinogen binding inhibitor which blocked the platelet aggregation. It is concluded that cytoskeletal proteins and GPIIb, IIIa are assembled each other and formed high molecular weight protein fraction or dimer formation during activation. In stimulated platelets these assembled cytoskeletal proteins containing GPIIb, IIIa were also found in Triton X-100 soluble fraction as a precursor of high molecular weight fraction in Triton X-100 insoluble fraction. The binding of fibrinogen to GPIIb-IIIa complex induce the linkage of GPIIb, IIIa to assembled cytoskeletal proteins.
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Yuping, Sun, Ma Xiaoli, Wang Yunshan, and Hu Anla. "Downregulation of AFP Protein Expression by RNAi Inhibits the Growth of AFP-Producing Gastric Adenocarcinoma Cell Line." In 2007 1st International Conference on Bioinformatics and Biomedical Engineering. IEEE, 2007. http://dx.doi.org/10.1109/icbbe.2007.15.

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Jakobs, K. H., P. Gierschik, and R. Grandt. "THE ROLE OF GTP-BINDING PROTEINS EXHIBITING GTPase ACTIVITY IN PLATELET ACTIVATION." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644773.

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Activation of platelets by agonists acting via cell surface-located receptors apparently involves as an early event in transmembrane signalling an interaction of the agonist-occupied receptor with a guanine nucleotide-binding regulatory protein (G-protein). The activated G-protein, then, transduces the information to the effector molecule, being responsible for the changes in intracellular second messengers. At least two changes in intracellular signal molecules are often found to be associated with platelet activation by agonists, i.e., increases in inositol trisphosphate and diacylglycerol levels caused by activation of a polyphosphoinositide-specific phospholipase C and decrease in cyclic AMP concentration caused by inhibition of adenylate cyclase.Both actions of platelet-activating agents apparently involve G-proteins as transducing elements. Generally, the function of a G-protein in signal transduction can be measured either by its ability to regulate the activity of the effector molecule (phospholipase C or adenylate cyclase) or the binding affinity of an agonist to its specific receptor or by the abitlity of the G-protein to bind and hydrolyze GTP or one of its analogs in response to agonist-activated receptors. Some platelet-activating agonists (e.g. thrombin) can cause both adenylate cyclase inhibition and phospholipase C activation, whereas others induce either inhibition of adenylate cyclase (e.g. α2-adrenoceptor agonists) or activation of phospholipase C (e.g. stable endoperoxide analogs) . It is not yet known whether the simultaneous activation of two signal transduction systems is due to activation of two separate G-proteins by one receptor, to two distinct receptors activating each a distinct G-protein or to activation of two effector molecules by one G-protein.For some of the G-proteins, rather specific compounds are available causing inactivation of their function. In comparison to Gs, the stimulatory G-protein of the adenylate cyclase system, the adenylate cyclase inhibitory Gi-protein is rather specifically inactivated by ADP-ribosylation of its a-subunit by pertussis toxin, “unfortunately” not acting in intact platelets, and by SH-group reactive agents such as N-ethylmaleimide and diamide, apparently also affecting the Giα-subunit. Both of these treatments completely block α2-adrenoceptor-induced GTPase stimulation and adenylate cyclase inhibition and also thrombin-induced inhibition of adenylate cyclase. In order to know whether the G-protein coupling receptors to phospholipase C is similar to or different from the Gi-protein, high affinity GTPase stimulation by agents known to activate phospholipase C was evaluated in platelet membranes. The data obtained indicated that GTPase stimulation by agents causing both adenylate cyclase inhibition and phospholipase C activation is reduced, but only partially, by the above mentioned Gi-inactivating agents, while stimulation of GTPase by agents stimulating only phospholipase C is not affected by these treatments. These data suggested that the G-protein regulating phospholipase C activity in platelet membranes is different from the Gi-protein and may also not be a substrate for pertussis toxin. Measuring thrombin stimulation of inositol phosphate and diacylglycerol formation in saponin-permeabilized platelets, apparently contradictory data were reported after pertussis toxin treatment, being without effect or causing even an increase in thrombin stimulation of inositol phosphate formation (Lapetina: BBA 884, 219, 1986) or being inhibitory to thrombin stimulation of diacylglycerol formation (Brass et al.: JBC 261, 16838, 1986). These data indicate that the nature of the phospholipase C-related G-protein(s) is not yet defined and that their elucidation requires more specific tools as well as purification and reconstitution experiments. Preliminary data suggest that some antibiotics may serve as useful tools to characterize the phospho-lipase-related G-proteins. The possible role of G-protein phosphorylation by intracellular signal molecule-activated protein kinases in attenuation of signal transduction in platelets will be discussed.
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Ma, Liang, Meixiang Xu, and Andres F. Oberhauser. "Nanoscale Analysis of the Effect of Pathogenic Mutations on Polycystin-1." In ASME 2010 First Global Congress on NanoEngineering for Medicine and Biology. ASMEDC, 2010. http://dx.doi.org/10.1115/nemb2010-13093.

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The activity of proteins and their complexes often involves the conversion of chemical energy (stored or supplied) into mechanical work through conformational changes. Mechanical forces are also crucial for the regulation of the structure and function of cells and tissues. Thus, the shape of eukaryotic cells is the result of cycles of mechano-sensing, mechano-transduction, and mechano-response. Recently developed single-molecule atomic force microscopy (AFM) techniques can be used to manipulate single molecules, both in real time and under physiological conditions, and are ideally suited to directly quantify the forces involved in both intra- and intermolecular protein interactions. In combination with molecular biology and computer simulations, these techniques have been applied to characterize the unfolding and refolding reactions in a variety of proteins, such as titin (an elastic mechano-sensing protein found in muscle) and polycystin-1 (PC1, a mechanosensor found in the kidney).
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Gralnick, H. R., L. M. Magurder, K. Hansmann, et al. "THE SURFACE EXPRESSION OF ALPHA GRANULE PROTEINS FOLLOWING THROMBIN STIMULATION." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643859.

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We have studied the platelet glycoproteins (GP) GPIb and the GPIIb/IIIa and the expression of alpha granule proteins (AGP) on the platelet (P) surface following thrombin (T) stimulation. The platelets were separated from plasma proteins on a arabinogalactan gradient. The P were stimulated with purified alpha T 0.1u/108p. Either monospecific polyclonal or murine monoclonal antibodies were used to detect the P glycoprotein and AGP. The platelets were analyzed on an EPICS V Flow Cytometer. Resting P had small amounts of AGP (2-8%) present on their surface. Within 1-3 min. after T stimulation significantly increased amounts of PF4 (26%) vWf (8%) Ig (10%) and the 140 kD alpha granule membrane (70%) were present on the P surface. The peak expression of all the AGP occurred within 5 mins. The 140 kD activation protein remained stable over 3-60 mins, in contrast the PF4 and the vWf expression peaked at 5 mins. and then decreased to near baseline levels. The GPIb and GPIIb/IIIa showed different patterns after activation. The GPIb intensity and number of positive cells decreased over time, while the GPIIb/ IIIa increased in flourescent intensity and the number of positive cells. These studies indicate that T stimulation of AGP on the P surface. vWf and P4 have a transient appearance on the P surface while Ig and the 140 kD activation protein both appear to become stable components of the P plasma membrane. This technique of detecting platelet activation is a specific, sensitive, and rapid method.
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Gray, R. J., Larry J. Kim, Lawrence J. DeLucas, Ali Arabshahi, and W. William Wilson. "Dynamic temperature-induced crystallization of proteins." In AIP Conference Proceedings Volume 387. ASCE, 1997. http://dx.doi.org/10.1063/1.52068.

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Ehrenberg, Anders. "Proteins, dynamics solutes with solvent properties." In AIP Conference Proceedings Volume 180. AIP, 1988. http://dx.doi.org/10.1063/1.37870.

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Discher, Dennis E., and Colin Johnson. "Alternative Splicing for Mechanical Resilience: The Softening Effect of Filamin’s Hinge." In ASME 2007 Summer Bioengineering Conference. American Society of Mechanical Engineers, 2007. http://dx.doi.org/10.1115/sbc2007-176751.

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Alternative splicing within proteins is common but not well understood in its influence on protein structure and stability. Filamins are ubiquitous actin-crosslinking proteins with two dozen Immunolgobulin (Ig) repeats and one alternatively-spliced ‘hinge’ that has been hypothesized to add flexibility. The hinge is also predicted to perturb folding. The molecular mechanics of filamins are probed here by AFM-forced extension, with a particular focus on the ∼30 aa hinge between repeats R15 and R16. After re-examining full-length filamin to clarify the single molecule limit for AFM experiments on long chains, short concatemers of (R15-R16)m and (R15-hinge-R16)m were studied by both AFM and solution structural methods. AFM shows that the hinged isoform extends and unfolds at smaller forces (60 pN) than the hinge-less form (80 pN), implying that the alternative splicing introduces a random coil that softens both adjacent domains. Circular Dichroism confirms that the hinge is a random coil, and thermal unfolding in solution suggests a weak destabilization by the hinge. Together with the rate-dependence of forced extension in AFM, the results reveal added resilience as the unfolding transition shifts to longer lengths upon insertion of the alternatively spliced hinge.
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Reports on the topic "Proteïna AFP"

1

Zhu, Xiaoyang, and Tim P. Lodge. Controlling Protein Conformation & Activities on Block-Copolymer Nanopatterns. Defense Technical Information Center, 2009. http://dx.doi.org/10.21236/ada520626.

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Weiss, Shimon. Biochemical and Physiological Characterization: Development & Apply Optical Methods for Charaterizing Biochemical Protein-Protein Interactions in MR-1. Office of Scientific and Technical Information (OSTI), 2006. http://dx.doi.org/10.2172/890585.

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Bednarek, Sebastian, Y. Role of AtCDC48 & the AtCDC48 Regulatory Protein Family, PUX, in Plant Cell Morphogenesis. Office of Scientific and Technical Information (OSTI), 2009. http://dx.doi.org/10.2172/977066.

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Imagawa, Walter T. Mechanisms of Altered Control of Proliferation by Cyclic Amp/Protein Kinase A During Mammary Tumor Progression. Defense Technical Information Center, 1999. http://dx.doi.org/10.21236/ada374059.

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LiWang, Andy. HBCU Equipment for AFOSR Project 13RSL012: The Mechanism by which ADP Regulates the Structure and Function of the Protein KaiC. Defense Technical Information Center, 2015. http://dx.doi.org/10.21236/ad1001100.

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Siuzdak, Gary. Ecosystems and Networks Integrated with Genes and Molecular Assemblies (ENIGMA): Component 5: Imaging Protein Conformations, Shapes & Assemblies in Solution & Administration project (Final Scientific/Technical Report, Subcontract No. 6974584). Office of Scientific and Technical Information (OSTI), 2021. http://dx.doi.org/10.2172/1797991.

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Stanley J. Opella. Structural Biology of The sequestration & Transport of Heavy Metal Toxins: NMR Structure Determination of Proteins Containing the CYS-X-Y-Metal Binding Motif. Office of Scientific and Technical Information (OSTI), 2004. http://dx.doi.org/10.2172/822065.

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Pawlowski, David R. Automated Sample Preparation (ASP): Development of a Rapid Method to Sequentially Isolate Nucleic Acids and Protein from Any Sample Type by a Cartridge-Based System. Defense Technical Information Center, 2013. http://dx.doi.org/10.21236/ada608052.

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