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）：单个 β-N-乙酰基-D-葡萄糖胺部分 (O-GlcNAc) 对核蛋白和细胞质蛋白的丝氨酸和苏氨酸进行 O-糖基化是一种普遍存在的高度动态且波动的翻译后修饰对细胞刺激作出反应。 O-GlcNAc 经常与蛋白质磷酸化竞争，这两种修饰在信号传导、转录以及癌基因和抑癌基因功能的调节中具有广泛的串扰。糖尿病是该应用的生物驱动因素，它是一种疾病的典型例子，其中胰岛素抵抗水平的增加、胰岛素抗凋亡作用的抑制、循环 O-GlcNAc 的改变已知会破坏正常信号传导，并且已被证实与此相关。诱导脂肪细胞因子水平、糖异生失调以及胰岛素基因转录的调节。我们将利用一种新的免疫原策略来开发针对三种蛋白质上四个 O-GlcNAc 修饰位点的位点特异性 O-GlcNac 抗体，这些蛋白质在糖尿病信号传导抑制中发挥作用。因此，如果我们成功，除了为此策略提供测试系统之外，这项初步研究中产生的单克隆抗体还将对糖尿病研究产生直接影响。我们预测，在第一阶段结束时，我们将展示一种策略，使 GlycoScientific 能够以已知且可控的成本生产各种 O-GlcNAc 位点特异性 MAb。我们认为这些将对疾病研究产生深远的影响。公共卫生相关性：单个 β-N-乙酰基-D-葡萄糖胺部分 (O-GlcNAc) 对核蛋白和细胞质蛋白的丝氨酸和苏氨酸进行 O-糖基化是一种普遍存在的翻译后修饰，它是高度动态的，并且会根据细胞刺激。 O-GlcNAc 经常与蛋白质磷酸化竞争，这两种修饰在信号传导、转录以及癌基因和抑癌基因功能的调节中存在广泛的串扰。糖尿病是该应用的生物驱动因素，它是一种疾病的典型例子，其中胰岛素抵抗水平的增加、胰岛素抗凋亡作用的抑制、循环 O-GlcNAc 的改变已知会破坏正常信号传导，并且已被证实与此相关。诱导脂肪细胞因子水平、糖异生失调以及胰岛素基因转录的调节。我们将利用一种新的免疫原策略来开发针对三种蛋白质上四个 O-GlcNAc 修饰位点的位点特异性 O-GlcNac 抗体，这些蛋白质在糖尿病信号传导抑制中发挥作用。因此，如果我们成功，除了为此策略提供测试系统之外，这项初步研究中产生的单克隆抗体还将对糖尿病研究产生直接影响。