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（通过应用提供）：“使用合成蛋白化学对O-GLCNAC修饰的功能分析” O-GLCNAC修饰（O-GLCNACYLATION）是一种动态蛋白质修饰，是哺乳动物胚胎发育绝对必需的，并且在包括疾病，包括糖尿病，神经膜，神经降解和癌症中。尽管大约1000种潜在蛋白是通过O-GLCNAC修饰的，但这些修饰对蛋白质功能的绝大多数影响是完全未知的。这种批判性缺乏知识的存在是因为传统方法是对特定地点O-Glcnacylation事件的研究确定性的。我们的研究计划的长期目标是了解O-Glcnacylation对蛋白质的后果，这是人类疾病关键的蛋白质。该应用的目标是制定蛋白质工程策略，这些策略可以独特地产生具有特定位点的O-GLCNAC修饰的蛋白质，并采用这些方法来了解O-Glcnacylation对蛋白A-核蛋白的影响，这是蛋白质核酸蛋白，这是帕克森病中骨料蛋白的蛋白质。我们的初步研究表明，可以使用合成化学制备同质的O-Glcnacylated蛋白。此外，我们已经使用合成蛋白化学来证明O-Glcnacylation会阻止A-核蛋白聚集。在这些初步研究的指导下，我们将：1）继续开发一般的合成形式，以制备O-Glcnacypated蛋白，2）研究O-Glcnacylation通过该机制阻止A-核蛋白聚集并确定O-Glcnacylation对细胞毒性的影响。这些研究很重要，因为O-Glcnacylation的影响几乎是完全未知的。此外，阻断A-核蛋白聚集是帕金森氏病的关键潜在理论策略。我们的方法也具有创新性，因为它使O-Glcnacylation的影响能够直接在A中进行测试特定于网站的方式，将来可以应用于其他关键蛋白质。