Proteomics Approaches to Protein Turnover

蛋白质周转的蛋白质组学方法

基本信息

批准号：
9027548
负责人：
JUNMIN PENG
金额：
$ 34.41万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-01-01 至 2019-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9027548
关键词：
Affect Alzheimer&apos s Disease Alzheimer&apos s disease model Amines Amino Acids Animal Disease Models Animals Antibodies Autophagocytosis Binding Biological Assay Cell Culture Techniques Cell Line Cells Complex Defect Development Digestion Eukaryotic Cell Event Functional disorder Genome Goals Label Lead Link Lysine Lysosomes Mammalian Cell Mass Spectrum Analysis Measurement Measures Mediating Methods Modification Molecular Molecular Biology Monitor Mus Pathogenesis Pathway interactions Physiologic pulse Play Polymers Polyubiquitin Protein Analysis Protein Biosynthesis Protein Dynamics Proteins Proteome Proteomics Protocols documentation Research Resolution Role Saccharomycetales Signal Transduction Specificity Stable Isotope Labeling Structure System Technology Trypsin Ubiquitin Ubiquitination Work Yeasts amino group base carbohydrate structure complex biological systems genetic information human disease improved innovation monomer mouse model multicatalytic endopeptidase complex novel protein degradation public health relevance tool transcriptome yeast genetics

项目摘要

DESCRIPTION (provided by applicant): The goal of this project is to develop proteomics technologies for the complete measurement of protein turnover in cells, focusing on the analysis of protein abundance, ubiquitination, synthetic rate, and degradation rate. According to the central dogma of molecular biology, genetic information flows from the genome to the transcriptome to the proteome. Theoretically, protein dynamics in a cellular system yields a steady state in which the level of a protein depends on its preexisting concentration, synthetic rate, and degradation rate. Protein degradation in eukaryotic cells is mainly mediated by the ubiquitin (Ub)-proteasome system and the autophagy-lysosome pathway. Ubiquitin, a small protein of 76 amino acids, modifies thousands of proteins as multifunctional signals for proteasomal degradation and other downstream events. Ubiquitin is conjugated to substrates in the form of monomers and polymers. In this application, we propose to develop novel mass spectrometry (MS)-based methods to profile protein turnover, analyze ubiquitinated proteome and ubiquitin chain structures, and study how protein turnover is affected under pathophysiological conditions. Our four specific aims are to (i) develop a 20-plex integrated MS approach for measuring proteome turnover; (ii) develop middle-down MS methods for analyzing polyubiquitin chain structures; (iii) study the function of diverse polyubiquitin chains in yeast ad mammalian cells; and (iv) investigate protein turnover alterations in mouse models of human disease. Protein turnover and ubiquitination are fundamental regulatory events, contributing to the pathogenesis of human disease. The research will lead to the development of novel MS technologies for studying protein turnover and new understanding of polyubiquitin chain function, as well as its involvement in human disease.

描述（由适用提供）：该项目的目的是开发蛋白质组学技术，以完全测量细胞中的蛋白质更新，重点是分析蛋白质丰度，泛素化，合成率和降解率。根据分子生物学的中心教条，遗传信息从基因组到转录组到蛋白质。理论，细胞系统中的蛋白质动力学产生稳态，在这种稳态中，蛋白质的水平取决于其先前存在的浓度，合成速率和降解速率。真核细胞中的蛋白质降解主要是由泛素（UB） - 培训体系统和自噬 - 溶质体途径介导的。泛素是一种76个氨基酸的小蛋白，将数千种蛋白质作为蛋白酶体降解和其他下游事件的多功能信号修饰。泛素以单体和聚合物的形式与底物结合。在此应用中，我们建议开发新的质谱法（MS）的方法来介绍蛋白质更新，分析泛素化蛋白质组和泛素链结构，并研究在病理生理条件下蛋白质更新如何影响蛋白质。我们的四个具体目的是（i）开发一种20次综合MS方法来测量蛋白质组更新；（ii）开发用于分析的多泛素链结构的中 - 向下MS方法；（iii）研究酵母AD哺乳动物细胞中潜水多泛素链的功能；（iv）研究人类疾病小鼠模型的蛋白质周转改变。蛋白质更新和泛素化是基本的调节事件，导致人类疾病的发病机理。这项研究将导致开发新型MS技术，用于研究蛋白质更新和对多泛素链功能的新理解及其参与人类疾病。