Site-specific O-GlcNAc MAbs: New Tools for Glycoproteomics

位点特异性 O-GlcNAc MAb：糖蛋白组学的新工具

基本信息

批准号：
8395145
负责人：
RON ORLANDO
金额：
$ 32.27万
依托单位：
GLYCOSCIENTIFIC, LLC
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-17 至 2014-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8395145
关键词：
Acetylglucosamine Affinity Antibodies Antigens Apoptotic Autoantigens Basic Science Biological Carbohydrates Cell physiology Complex Cytoplasmic Protein Detection Diabetes Mellitus Disease Enzymes Epitopes Genetic Transcription Gluconeogenesis Glycopeptides Harvest Heart Human Hybridomas Immune system Immunization Injectable Insulin Insulin Resistance Lead Letters Libraries Link Modification Monoclonal Antibodies Mus Nuclear Proteins Phase Phospho-Specific Antibodies Phosphorylation Play Polysaccharides Post-Translational Protein Processing Preparation Process Production Protein-Carbohydrate Interaction Proteins Reagent Regulation Research Research Personnel Resources Role Scientist Screening procedure Serine Signal Transduction Site Specificity Stimulus Synthetic Vaccines System Testing Threonine Tumor Suppressor Genes UDP-N-acetylglucosamine-peptide beta-N-acetylglucosaminyltransferase Universities Western Blotting biological systems candidate selection cost glycosylation immunogenicity medical schools peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase peptide structure professor protein aminoacid sequence response tool

项目摘要

DESCRIPTION (provided by applicant): O-glycosylation of serine and threonine of nuclear and cytoplasmic proteins by a single beta-N- acetyl-D-glucosamine moiety (O-GlcNAc) is a ubiquitous post-translational modification that is highly dynamic and fluctuates in response to cellular stimuli. O-GlcNAc often competes with protein phosphorylation, and these two modifications have extensive crosstalk in the regulation of signaling, transcription, and the functions of oncogenes and tumor suppressors. Diabetes, the biological driver for this application, is a prime example of a disease where increased levels of insulin resistance, inhibition of the anti-apoptotic action of insulin, alteration of circulating O-GlcNAc is known to disrupt normal signaling, and has been associated with the induction of adipocytokine levels, deregulation of gluconeogenesis, and modulation of insulin gene transcription. We will utilize a new immunogen strategy to develop site-specific O-GlcNac antibodies to four sites of O-GlcNAc modification on three proteins that play a role in signaling suppression in diabetes. Consequently, if we are successful, the mAbs generated in this initial study will have an immediate impact on diabetes research in addition to providing a test system for this strategy. We predict that at the end of phase 1, we will have demonstrated a strategy that will allow GlycoScientific to produce a wide range of O-GlcNAc site-specific MAbs, at a known and contained cost. We feel that these will have far reaching implications in disease research. PUBLIC HEALTH RELEVANCE: O-glycosylation of serine and threonine of nuclear and cytoplasmic proteins by a single beta-N- acetyl-D-glucosamine moiety (O-GlcNAc) is a ubiquitous post-translational modification that is highly dynamic and fluctuates in response to cellular stimuli. O-GlcNAc often competes with protein phosphorylation, and these two modifications have extensive crosstalk in the regulation of signaling, transcription, and the functions of oncogenes and tumor suppressors. Diabetes, the biological driver for this application, is a prime example of a disease where increased levels of insulin resistance, inhibition of the anti-apoptotic action of insulin, alteration of circulating O-GlcNAc is known to disrupt normal signaling, and has been associated with the induction of adipocytokine levels, deregulation of gluconeogenesis, and modulation of insulin gene transcription. We will utilize a new immunogen strategy to develop site-specific O-GlcNac antibodies to four sites of O-GlcNAc modification on three proteins that play a role in signaling suppression in diabetes. Consequently, if we are successful, the mAbs generated in this initial study will have an immediate impact on diabetes research in addition to providing a test system for this strategy.

描述（由申请人提供）：通过单个β-N-乙酰基-D-葡萄糖胺基部分（O-GLCNAC）对核和细胞质蛋白的丝氨酸和苏氨酸的O-糖基化是一种高度动态的，并且对细胞刺激的反应是高度动力学的，是一种无处不在的后翻译后修饰。 O-GLCNAC经常与蛋白质磷酸化竞争，这两种修饰在信号，转录和致癌基因和肿瘤抑制器的功能的调节中具有广泛的串扰。 Diabetes, the biological driver for this application, is a prime example of a disease where increased levels of insulin resistance, inhibition of the anti-apoptotic action of insulin, alteration of circulating O-GlcNAc is known to disrupt normal signaling, and has been associated with the induction of adipocytokine levels, deregulation of gluconeogenesis, and modulation of insulin gene transcription. 我们将利用一种新的免疫原策略来开发特异性的O-GLCNAC抗体，以对三种蛋白质的O-GLCNAC修饰位点开发三种蛋白质，这些蛋白质在糖尿病的信号抑制中起作用。因此，如果我们取得成功，则在这项初步研究中产生的mABS除了为该策略提供测试系统外，还将对糖尿病研究产生直接影响。我们预测，在第1阶段结束时，我们将展示一种策略，该策略将允许糖科学以已知和包含的成本产生各种O-GLCNAC特定于特异性的单位MAB。我们认为这些对疾病研究具有很大的影响。公共卫生相关性：单个β-N-乙酰基-D-葡萄糖苷部分（O-GLCNAC）对核和细胞质蛋白的丝氨酸和苏氨酸的O-糖基化是一种无处不在的后翻译后修饰，是对细胞刺激的反应强烈的动态和波动。 O-GLCNAC经常与蛋白质磷酸化竞争，这两种修饰在信号，转录和致癌基因和肿瘤抑制器的功能的调节中具有广泛的串扰。 Diabetes, the biological driver for this application, is a prime example of a disease where increased levels of insulin resistance, inhibition of the anti-apoptotic action of insulin, alteration of circulating O-GlcNAc is known to disrupt normal signaling, and has been associated with the induction of adipocytokine levels, deregulation of gluconeogenesis, and modulation of insulin gene transcription. 我们将利用一种新的免疫原策略来开发特异性的O-GLCNAC抗体，以对三种蛋白质的O-GLCNAC修饰位点开发三种蛋白质，这些蛋白质在糖尿病的信号抑制中起作用。因此，如果我们取得成功，则在这项初步研究中产生的mABS除了为该策略提供测试系统外，还将对糖尿病研究产生直接影响。