Functional Analysis of O-GlcNAc using Synthetic Protein Chemistry

使用合成蛋白质化学对 O-GlcNAc 进行功能分析

基本信息

批准号：
10671580
负责人：
Matthew Robert Pratt
金额：
$ 34.69万
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 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10671580
关键词：
Acetylation Affect Amyloid Animals Biochemical Biochemistry Biological Biology Biophysics Carbohydrates Cellular biology Charcot-Marie-Tooth Disease Chemicals Collaborations Cryoelectron Microscopy Data Development Diabetes Mellitus Diffusion Disease Embryonic Development Excision Functional disorder Funding Goals Grant HSPB1 gene Heat shock proteins Kinetics Knowledge Ligation Malignant Neoplasms Mammals Measures Methods Michigan Modification Molecular Molecular Chaperones Monosaccharides Mus Mutagenesis Mutation Nerve Degeneration Neurodegenerative Disorders Neurons Neuropathy O-GlcNAc transferase Parkinson Disease Pathogenicity Pathway interactions Patients Peripheral Nervous System Diseases Phosphorylation Play Point Mutation Polymorph Post-Translational Protein Processing Protein Chemistry Protein Dynamics Proteins Proteomics Publishing Recombinant Proteins Research Role Serine Serine/Threonine Phosphorylation Silicon Dioxide Site Structure Testing Therapeutic Threonine Toxic effect Western Blotting Work alpha synuclein amyloid formation amyloid structure biophysical properties dark matter experimental study glycosylation human disease in vivo monomer mutant prevent programs protein aggregation protein function public health relevance stereochemistry sugar synthetic protein unnatural amino acids

项目摘要

Modified Project Summary/Abstract (The abstract contained no specific references to the in vivo mouse experiments and therefore is unchanged): O-GlcNAc modification 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 1,000 proteins are modified by O-GlcNAc, the effects of the vast majority of these modifications on protein function are completely unknown. The long-term goal of our research program is to fill in these missing gaps by determining the biochemical consequences of O-GlcNAc on proteins that are key to human disease. To accomplish this goal, we use a combination of carbohydrate and synthetic protein chemistries to build O-GlcNAc modified proteins for subsequent biological experiments. This chemical approach is uniquely enabling, as it is currently the only way to generate homogeneous and site-specifically O-GlcNAc modified proteins. We have been very successful and have used synthetic proteins to determine that O-GlcNAc has a multifaceted role in preventing the amyloid aggregation of proteins in neurodegenerative diseases. Specifically, we have found that O-GlcNAc both directly inhibits the aggregation of amyloid forming proteins and activates the activity of certain small chaperones. In this proposal we will continue to build on these discoveries. In Aim 1, we will determine how O-GlcNAc inhibits the early stages of α-synuclein amyloid formation. In Aim 2, we will test whether O-GlcNAc alters the structure/toxicity relationships of α-synuclein amyloids. In Aim 3, we examine how O-GlcNAc alters the small heat shock protein interactome. Finally, in Aim 4, we will determine if O-GlcNAc can rescue the activity of mutant chaperones that cause Charcot-Marie-Tooth disease. At the conclusion of these independent aims, we will have further unravelled the mechanisms by which O-GlcNAc inhibits protein aggregation and provided critical data to support the ongoing efforts to target O-GlcNAc therapeutically.

修改的项目摘要/摘要（摘要中没有对体内小鼠实验的特定引用，因此没有变化）CNAC修饰是一种动态蛋白质修饰，是哺乳动物的胚胎发育绝对的，S，S，含糖尿病，神经变性生成和癌症的疾病。 O-GlcNAC对垂体进行了大约1,000个蛋白质的修饰，这些蛋白质功能的修饰是我们研究计划的长期目标是通过确定生物化学惯例的蛋白质来填补这些缺失的差距合成蛋白化学以构建O-GlCNAC修饰的蛋白质，以进行生物学实验，这是产生同质和特异性O-GLCNAC修饰蛋白的唯一方法。在神经退行性疾病中的产生。 O-GLCNAC抑制α-突触核蛋白淀粉样蛋白的thibits在AIM 2 ps的α-核蛋白淀粉样蛋白中。 O-GLCNAC可以在独立目标结束时挽救突变伴侣伴侣的作用靶向O-GLCNAC治疗。

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Understanding and exploiting the roles of O-GlcNAc in neurodegenerative diseases.

DOI：
10.1016/j.jbc.2023.105411
发表时间：
2023-12
期刊：
JOURNAL OF BIOLOGICAL CHEMISTRY
影响因子：
4.8
作者：
Pratt, Matthew R.;Vocadlo, David J.
通讯作者：
Vocadlo, David J.

The E3 ligase adapter cereblon targets the C-terminal cyclic imide degron.

DOI：
10.1038/s41586-022-05333-5
发表时间：
2022-10
期刊：
NATURE
影响因子：
64.8
作者：
Ichikawa, Saki;Flaxman, Hope A.;Xu, Wenqing;Vallavoju, Nandini;Lloyd, Hannah C.;Wang, Binyou;Shen, Dacheng;Pratt, Matthew R.;Woo, Christina M.
通讯作者：
Woo, Christina M.

Optimization of Chemoenzymatic Mass Tagging by Strain-Promoted Cycloaddition (SPAAC) for the Determination of O-GlcNAc Stoichiometry by Western Blotting.