Characterizing proteasome-substrate interactions by mass spectrometry proteomics

通过质谱蛋白质组学表征蛋白酶体-底物相互作用

基本信息

批准号：
10200097
负责人：
DAVID E. CLEMMER
金额：
$ 34.18万
依托单位：
TRUSTEES OF INDIANA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10200097
关键词：
Affinity Analytical Chemistry Attenuated Binding Biochemical Biochemical Genetics Biochemistry Bioinformatics Biological Biological Assay Cell physiology Cells Cellular biology Chemicals Collaborations Complex Data Data Analyses Development Discipline Disease Resistance Elements Genetic Geometry Hematologic Neoplasms Immune response In Vitro Indiana Ions Kinetics Ligands Maintenance Mass Spectrum Analysis Methodology Methods Modification Molecular Biology Molecular Structure Mutagenesis Neurodegenerative Disorders Peptides Physiological Polyubiquitin Process Proteasome Binding Proteasome Inhibitor Protein Engineering Proteins Proteome Proteomics Quality Control Reporting Research Research Personnel Role Signal Transduction Site Specificity Structure Substrate Interaction System Testing Texas Ubiquitin Ubiquitination Universities Yeasts austin crosslink data acquisition defined contribution detection sensitivity experience experimental study genetic regulatory protein healthy aging improved instrumentation multicatalytic endopeptidase complex novel polypeptide protein degradation proteostasis purge receptor therapeutic target yeast genetics

项目摘要

PROJECT SUMMARY This application features a close collaboration between bioanalytical (Dr. Reilly, Indiana University) and bio- chemistry (Dr. Matouschek, University of Texas, Austin) research groups aimed at improving our understanding of the ubiquitin-proteasome protein degradation system. The investigators contribute complementary expertise and experience in a variety of disciplines ranging from proteomics, protein bioinformatics, analytical chemistry and instrumentation, to biochemistry, molecular biology and cell biology. At Indiana University, cross-linking mass spectrometry will be combined with biochemical and genetic assays to characterize the interaction between the proteasome and its substrates to provide candidates for novel substrate receptors on the proteasome. At The University of Texas at Austin, the biological relevance of candidates will then be tested in biochemical pro- teasome assays and genetic yeast experiments. These assays will define the contribution of newly identified components to substrate recognition and degradation by the proteasome and elucidate the molecular and struc- tural origin of substrate selection and specificity. Overall, we believe that this proposal will result in significant advances in cross-linking methodology and in our understanding of how the proteasome recognizes and digests substrates.

项目摘要该应用程序具有生物分析（Reilly博士，印第安纳大学）和Bio-之间的密切合作化学（德克萨斯大学奥斯汀分校Matouschek博士）研究小组旨在提高我们的理解泛素 - 蛋白酶体蛋白质降解系统的。研究人员贡献了补充专业知识以及各种学科的经验，包括蛋白质组学，蛋白质生物信息学，分析化学和仪器，用于生物化学，分子生物学和细胞生物学。在印第安纳大学，交联质谱法将与生化和遗传测定法相结合，以表征蛋白酶体及其底物为蛋白酶体上新型底物受体提供候选。在德克萨斯大学奥斯汀分校，候选人的生物学相关性将在生化培训中进行测试 Teasome分析和遗传酵母实验。这些测定将定义新确定的贡献通过蛋白酶体进行底物识别和降解的成分，并阐明分子和结构底物选择和特异性的胞外起源。总体而言，我们认为该提案将在交联方法和我们的了解蛋白酶体如何识别和消化底物。