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DEFINING THE FUNCTION OF PROTEASOMAL DEUBIQUITINASES

定义蛋白酶体去泛素酶的功能

基本信息

批准号：
10221698
负责人：
ERIC Robert STRIETER
金额：
$ 36.23万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10221698
关键词：
26S proteasome ATP-Dependent Proteases Address Affect Architecture Attention Binding Biological Biological Assay CRISPR/Cas technology Cell Line Cells Cellular biology Cleaved cell Complex Crystallography Cyclic Peptides Data Deubiquitination Deuterium Development Disease Embryo Enzymes Failure Funding Generations Health Human Human Biology Hydrogen Kinetics Knowledge Label Light Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Modification Molecular Neurodegenerative Disorders Pathway interactions Peptide Hydrolases Phage Display Physiological Play Polymers Positioning Attribute Process Proteasome Binding Protein Chemistry Proteins Proteomics Research Role Signal Transduction Specificity Structure System Testing Translations Ubiquitin Work X-Ray Crystallography cellular targeting genome editing inhibitor/antagonist innovation insight interdisciplinary approach kidney cell multicatalytic endopeptidase complex novel novel therapeutics premature prevent protein degradation proteostasis small molecule therapeutic target tool

项目摘要

Project Summary The 26S proteasome is a massive, intricately regulated ATP-dependent protease that is responsible for the degradation of most cellular proteins. Failure of the proteasome to precisely regulate protein levels is a hallmark of many cancers and neurodegenerative diseases. Targeting proteins to the proteasome requires covalent modification with a polymeric (Ub) chain. Thus, it is perplexing that the proteasome actually houses enzymes (deubiquitinases/DUBs) responsible for removing Ub chains. While it has become clear that the intrinsic DUB, RPN11, promotes degrades by preventing premature deubiquitination of proteins not yet engaged with the proteasome, the roles of other proteasomal DUBs, e.g., UCH37/UCHL5, are poorly understood. In preliminary studies, we discovered that proteasome-bound UCH37 acts as a chain editor by removing branch points. This application proposes to elucidate how UCH37 selects branched Ub chains for editing, how this activity is integrated into the entire process of proteasomal degradation, and how it impacts the turnover of cellular proteins. In Aim 1, we will investigate the role of UCH37-mediated chain debranching during proteasomal degradation using distinct, purified human proteasome complexes and fluorescent, polyubiquitinated substrates. In Aim 2, we propose to identify cellular targets of proteasome-bound UCH37 using an innovative combination of quantitative proteomics, in-cell proximity labeling, and Ub middle-down mass spectrometry. In Aim 3, we focus on understanding the molecular basis of UCH37's specificity toward branched chains. Finally, in Aim 4, we will develop novel cyclic peptide inhibitors of UCH37 to facilitate efforts to dissect its function in any biological paradigm. The knowledge gained from this research will shed light on fundamental aspects of the ubiquitin proteasome system and pave the way for the development of new therapeutics that regulate proteasome function.

项目总结