Comprehensive proteomic profiling of the ubiquitin pathway in cancer

癌症中泛素通路的全面蛋白质组学分析

• 批准号: 7855709
• 负责人: DIETER A WOLF
• 金额: $ 92.31万
• 依托单位: SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7855709
• 关键词: Address; Adopted; Androgens; Biological Assay; Biological Markers; Biological Process; Biology; Cells; Data; Data Set; Databases; Development; Diagnostic Neoplasm Staging; Disease; Enzymes; Eukaryota; Foundations; Genomics; Goals; Human; Inflammation; Link; Lysine; Malignant Neoplasms; Malignant neoplasm of prostate; Mass Spectrum Analysis; Measurement; Methodology; Modeling; Names; Nerve Degeneration; Pathway interactions; Patients; Pharmacologic Substance; Phase; Phenotype; Post-Translational Protein Processing; Prostate; Proteins; Proteomics; Regulation; Research Personnel; Resistance; Resources; Screening procedure; Solid; Solutions; System; Technology; Therapeutic; Ubiquitin; Virus Diseases; base; cell growth regulation; functional genomics; human disease; immortalized cell; improved; novel; novel diagnostics; public health relevance; tool; tumor; tumor initiation; tumor progression; tumorigenic

DESCRIPTION (provided by applicant): The ubiquitin system of protein modification permeates every field in biology and is intimately linked to the most prevalent human diseases, including cancer, neurodegeneration, inflammation, and viral infections. With ~50 E2s and an estimated several hundred E3s, the ubiquitin system constitutes one of the most byzantine enzyme systems in eukaryotes. Whereas the basic principle of the E1-E2-E3 enzymatic cascade is now well established, the biological functions and regulation of ubiquitylation enzymes remain largely a mystery, mainly due to our inability to rapidly and comprehensively match ubiquitylation enzymes with their cellular substrates. As a solution to this predicament, which forestalls the entire ubiquitin field, a new activity-based protein capture and profiling technology, named SPASS (Solid Phase Assay for Ubiquitylation Substrate Screening) is proposed. SPASS simultaneously addresses three contemporary challenges: (1) It allows the unbiased identification of ubiquitylation substrates in an E2-specific manner. (2) Whereas structural information has proven unable to match the ~50 E2 with their hundreds of cooperating E3s, SPASS methodology provides this capability. (3) Lastly, the specific lysine residues that are modified are unknown for most substrates, a shortcoming that is equally addressed by the SPASS substrate capture and identification methodology. In the present project, SPASS will be applied to the profiling of ~15 E2s in a syngeneic tumor initiation and progression model of human prostate cancer. The main goal is to identify the substrates and cooperating E3 enzymes of these E2s, to pinpoint the substrate lysines that are modified, and to quantify differences in substrate utilization as prostate cells progress from an immortalized to a tumorigenic phenotype. The main deliverables will be a comprehensive platform and publicly available datasets that will provide cancer researchers and clinicians with a novel resource to link genomic and proteomic data with cancer progression. PUBLIC HEALTH RELEVANCE: The essential involvements of ubiquitylation enzymes (E2 and E3) in cellular regulation have placed them squarely at the center of prevalent human diseases, in particular cancer. Consequently, several pharmaceutical companies are already developing strategies for modulating ubiquitin system activity in a therapeutic context. A comprehensive understanding of the exact substrates and pathways controlled by ubiquitylation enzymes such as enabled by the studies proposed in this application will lay the foundation for harnessing the full potential of such efforts.

描述（由申请人提供）：蛋白质修饰的泛素系统渗透到生物学的每个领域，并与最普遍的人类疾病密切相关，包括癌症，神经变性，炎症和病毒感染。泛素系统含有约50个E2s和数百个E3s，是真核生物中最复杂的酶系统之一。尽管E1-E2-E3酶级联的基本原理已经确立，但泛素化酶的生物学功能和调控在很大程度上仍然是一个谜，这主要是因为我们无法快速全面地将泛素化酶与其细胞底物匹配起来。为了解决这一困扰整个泛素领域的困境，一种新的基于活性的蛋白质捕获和分析技术被提出，称为SPASS（固相法泛素化底物筛选）。SPASS同时解决了三个当代挑战：(1)它允许以e2特异性的方式公正地鉴定泛素化底物。(2)虽然结构信息已被证明无法将~50 E2与其数百个协同e3进行匹配，但SPASS方法提供了这种能力。(3)最后，对于大多数底物来说，被修饰的特定赖氨酸残基是未知的，SPASS底物捕获和鉴定方法同样解决了这一缺点。在本项目中，SPASS将应用于人类前列腺癌同基因肿瘤发生和发展模型中~15个E2s的谱分析。主要目标是鉴定这些E2s的底物和合作E3酶，确定被修饰的底物赖氨酸，并量化前列腺细胞从永生化表型向致瘤表型发展过程中底物利用的差异。主要成果将是一个全面的平台和公开可用的数据集，这将为癌症研究人员和临床医生提供一种新的资源，将基因组和蛋白质组学数据与癌症进展联系起来。