Functional Analysis of O-GlcNAc Modifications Using Synthetic Protein Chemistry

使用合成蛋白质化学对 O-GlcNAc 修饰进行功能分析

基本信息

批准号：
9321152
负责人：
Matthew Robert Pratt
金额：
$ 32.59万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-01 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9321152
关键词：
Affect Alzheimer&apos s Disease Biochemical Biological Assay Biophysics Carbon Cell Culture Techniques Cells Chemicals Complex Data Development Diabetes Mellitus Disease Embryonic Development Enzymes Event Functional disorder Future Generations Glucosamine Goals Health In Vitro Individual Insecta Knowledge Letters Lewy Bodies Link Literature Malignant Neoplasms Mammals Methods Mitochondrial Proteins Modeling Modification Molecular Monosaccharides Nerve Degeneration Neurons Nuclear Proteins Parkinson Disease Parkinson&apos s Dementia Physiological Positioning Attribute Post-Translational Protein Processing Preparation Protein Chemistry Protein Engineering Proteins Publications Reaction Reagent Research Route Science Site Synthesis Chemistry Testing Therapeutic Toxic effect Ubiquitination alpha synuclein analog biophysical properties collaborative environment experimental study extracellular human disease in vivo innovation mouse model peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase prevent prion-like programs protein aggregate protein function public health relevance synthetic protein tau Proteins transmission process

项目摘要

DESCRIPTION (provided by applicant): "Functional Analysis of O-GlcNAc Modifications using Synthetic Protein Chemistry" O-GlcNAc modification (O-GlcNAcylation) is a dynamic protein-modification that is absolutely required for embryonic development in mammals, and is misregulated in diseases, including diabetes, neurodegeneration and cancer. Although approximately 1000 potential proteins are modified by O-GlcNAc, the effects of the vast majority of these modifications on protein function are completely unknown. This critical lack of knowledge exists in-part because traditional methods are deficient for the study of site-specific O-GlcNAcylation events. The long-term goal of our research program is to understand the consequences of O-GlcNAcylation on proteins that are key to human disease. The objectives of this application are to develop protein engineering strategies that uniquely enable the generation of proteins with site-specific O-GlcNAc modifications and to apply these methods to understand the effects of O-GlcNAcylation on the protein a-synuclein, the aggregation-prone protein in Parkinson's disease. Our preliminary studies demonstrate that homogeneously O-GlcNAcylated proteins can be prepared using synthetic chemistry. Furthermore, we have used synthetic protein chemistry to demonstrate that O-GlcNAcylation blocks a-synuclein aggregation. Guided by these preliminary studies, we will: 1) continue to develop general synthetic-strategies for the preparation of O-GlcNAcylated proteins, 2) investigate the molecular mechanism by which O-GlcNAcylation blocks a-synuclein aggregation and 3) determine the effects of O-GlcNAcylation on the cellular toxicity of a-synuclein. These studies are significant, as the effects of O-GlcNAcylation are almost completely unknown. Additionally, blocking a-synuclein aggregation is a key potential therapeutic strategy in Parkinson's disease. Our approach is also innovative as it enables the effects of O-GlcNAcylation to be directly tested in a site-specific fashion and can be applied to other critical proteins in the future.

描述（由申请人提供）：“使用合成蛋白质化学对 O-GlcNAc 修饰进行功能分析”O-GlcNAc 修饰（O-GlcNAc 修饰）是一种动态蛋白质修饰，是哺乳动物胚胎发育所绝对必需的，并且在疾病中受到错误调节，包括糖尿病、神经退行性变和癌症，尽管 O-GlcNAc 修饰了大约 1000 种潜在蛋白质，但其中绝大多数修饰的影响这种严重缺乏知识的部分原因是传统方法不足以研究位点特异性 O-GlcNAc 酰化事件。我们研究计划的长期目标是了解 O-GlcNAc 酰化的后果。该应用的目标是开发蛋白质工程策略，能够独特地生成具有位点特异性 O-GlcNAc 修饰的蛋白质，并应用这些方法来了解 O-GlcNAc 酰化对人类疾病的影响。蛋白质α-突触核蛋白是帕金森病中易于聚集的蛋白质，我们的初步研究表明，可以使用合成化学来制备均一的 O-GlcNA 酰化蛋白。此外，我们还使用合成蛋白质化学来证明 O-GlcNA 酰化可阻止 α-突触核蛋白聚集。在这些初步研究的指导下，我们将：1）继续开发制备 O-GlcNA 酰化蛋白的通用合成策略，2）通过以下方法研究分子机制：其中O-GlcNAcylation阻断α-突触核蛋白聚集，并且3)确定O-GlcNAcylation对α-突触核蛋白的细胞毒性的影响这些研究是重要的，因为O-GlcNAcylation的影响几乎完全未知。突触核蛋白聚集是帕金森病的一种关键的潜在治疗策略，我们的方法也是创新的，因为它可以直接测试 O-GlcNAcylation 的效果。位点特异性时尚，将来可以应用于其他关键蛋白质。