Analysis of the VHL-HIF hydroxylase pathway

VHL-HIF羟化酶途径分析

• 批准号: 7669079
• 负责人: YUZURU SHIIO
• 金额: $ 22.19万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-25 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7669079
• 关键词: Affinity Chromatography; Apoptosis; Bacteriophages; Binding; Binding Proteins; Biochemical; Biogenesis; Biological; Biological Phenomena; Biological Process; Biology; Cells; Complex; EP300 gene; Hydroxylation; Hypoxia Inducible Factor; Inherited; Iron; Iron Chelation; Isotopically-Coded Affinity Tagging; MYBBP1A gene; Mediating; Mediator of activation protein; Methods; Mixed Function Oxygenases; Molecular Profiling; Mutation; Oncogene Proteins; Oncogenes; Oxidative Phosphorylation; Oxygen; Pathway interactions; Pheochromocytoma; Play; Post-Translational Protein Processing; Proline; Proteins; Proteomics; Renal Cell Carcinoma; Role; Screening procedure; Technology; Transcription Coactivator; Tumor Angiogenesis; Tumor Suppression; Tumor Suppressor Genes; Ubiquitin; Ubiquitination; VHL mutation; anticancer research; base; expression cloning; follow-up; hemangioblastoma; interest; multicatalytic endopeptidase complex; novel; novel strategies; protein complex; protein expression; tumor; tumorigenesis; ubiquitin ligase

DESCRIPTION (provided by applicant): Protein modification with ubiquitin plays key regulatory roles in a wide variety of biological phenomena including tumorigenesis. Several oncoproteins and anti-oncoproteins display ubiquitin ligase activity and identification of their substrates is one of the central themes in cancer research. This application is directed toward understanding the biological roles of a tumor-suppressing ubiquitin ligase, VHL. VHL is currently best understood as a negative regulator of hypoxia inducible factor (HIF). In the presence of oxygen and iron, specific proline residues in HIF are hydroxylated and these hydroxylated prolines are recognized by VHL, which results in ubiquitination and degradation of HIF. VHL mutation is associated with hereditary and sporadic renal cell carcinomas, hemangioblastomas, and pheochromocytomas. Several lines of evidence suggest that, in addition to HIF, there are unidentified substrates for VHL which play critical roles in tumor suppression. A major barrier to understanding the VHL functions (as well as the functions of other ubiquitin ligases) has been the difficulty in pinpointing its ubiquitination substrates. Employing a novel quantitative proteomics technology, ICAT (isotope-coded affinity tag), we developed an approach to systematically identify the substrates of ubiquitin ligases and using this method, we identified a novel substrate for VHL, which carries the character of a key mediator of VHL functions. Therefore the aims of this application are to analyze the mechanism and biological consequence of ubiquitination of this substrate and to conduct further proteomic screens to identify additional substrates for VHL. These studies should provide a better understanding of the biological functions of VHL and advance the nascent field of proteomics-based biology.

描述（由申请人提供）：泛素蛋白修饰在包括肿瘤发生在内的多种生物现象中发挥着关键的调节作用。几种癌蛋白和抗癌蛋白表现出泛素连接酶活性，其底物的鉴定是癌症研究的中心主题之一。该应用旨在了解肿瘤抑制泛素连接酶 VHL 的生物学作用。 VHL 目前最好被理解为缺氧诱导因子 (HIF) 的负调节因子。在氧和铁存在的情况下，HIF 中的特定脯氨酸残基被羟基化，这些羟基化的脯氨酸被 VHL 识别，从而导致 HIF 泛素化和降解。 VHL 突变与遗传性和散发性肾细胞癌、血管母细胞瘤和嗜铬细胞瘤有关。多项证据表明，除了 HIF 之外，还有未鉴定的 VHL 底物，它们在肿瘤抑制中发挥着关键作用。了解 VHL 功能（以及其他泛素连接酶的功能）的一个主要障碍是难以精确定位其泛素化底物。采用新型定量蛋白质组学技术 ICAT（同位素编码亲和标签），我们开发了一种系统鉴定泛素连接酶底物的方法，并使用该方法，我们鉴定了 VHL 的一种新型底物，它具有泛素连接酶关键介质的特征。 VHL 功能。因此，本申请的目的是分析该底物泛素化的机制和生物学后果，并进行进一步的蛋白质组筛选以鉴定 VHL 的其他底物。这些研究应该可以更好地理解 VHL 的生物学功能，并推动基于蛋白质组学的生物学的新兴领域的发展。