Molecular Regulation of Human iNOS by Phosphorylation

人 iNOS 磷酸化的分子调控

• 批准号: 6934576
• 负责人: YI XU
• 金额: $ 5.75万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6934576
• 关键词: biosynthesis; cofactor; enzyme activity; enzyme substrate; human tissue; immunoprecipitation; ligase; mass spectrometry; nitric oxide; nitric oxide synthase; phosphorylation; postdoctoral investigator; posttranslational modifications; protein degradation; protein purification; respiratory epithelium; tissue /cell culture; ubiquitin

DESCRIPTION (provided by applicant): Overproduction of nitric oxide (NO) by inducible nitric oxide synthase (iNOS) has been implicated in the pathogenesis of several diseases including airway lung inflammation. The overall objective of this proposal is to understand the posttranslational mechanisms of cellular turnover of iNOS and thus lay the groundwork for future studies aiming at controlling NO synthesis. Our preliminary data suggest that iNOS is subject to regulation by phosphorylation precedes its ubiquitination. We propose to test the following hypotheses: A) The specific ubiquitin ligase that links ubiquitin to iNOS binds preferentially to phosphorylated iNOS. Thus phosphorylation of specific iNOS residue(s) is critical for iNOS ubiquitination and degraqdation. B) Cells maintain a tight control over NO synthesis by maintaining a robust rate of iNOS turnover that is modulated by cellular factors. To test these hypotheses we propose studies with the following Specific Aims; Aim 1: Elucidation of the role of phosphorylation in iNOS turnover and determining specific phosporylated residues; Aim 2: Characterization of the rate of iNOS turnover and cellular factors that modulate it. The rational for the proposed studies is that once these mechanisms are understood, therapeutic strategies can be designed to alter these pathways and modulate iNOS turnover.

描述（由申请人提供）：诱导型一氧化氮合酶（INOS）对一氧化氮（NO）的生产过量与包括气道肺炎症在内的多种疾病的发病机理有关。 该提案的总体目的是了解iNOS细胞周转的翻译后机制，从而为未来的研究奠定了旨在控制无综合的研究的基础。 我们的初步数据表明，iNOS受磷酸化在其泛素化之前受到调节。 我们建议检验以下假设：a）将泛素与iNOS联系起来的特异性泛素连接酶优先结合与磷酸化的iNOS。 因此，特异性iNOS残基的磷酸化对于ionos泛素化和降解至关重要。 b）细胞通过维持由细胞因子调节的iNOS周转率的稳健速率来维持对NO合成的严格控制。 为了检验这些假设，我们提出了以下特定目标的研究；目的1：阐明磷酸化在iNOS周转率中的作用并确定特定的芬比利残基； AIM 2：表征调节iNOS更新和细胞因子的速率。 拟议的研究的理性是，一旦理解了这些机制，就可以设计治疗策略来改变这些途径并调节iNOS转换。