Molecular Mechanisms of iNOS Degradation

描述（由申请人提供）：候选人是一个最近独立的医生，科学家谁表现出强烈的承诺，生物医学研究。他的长期职业目标是在气道炎症的分子机制方面发展强大的研究计划。他的近期研究目标是了解气道细胞中诱导型一氧化氮合酶（iNOS）调节NO合成的分子机制，并设计控制方法。成功的R 01申请使候选人准备继续他目前的研究，启动新的项目，获得新的技能和建立新的合作。需要K 02奖来减少候选人当前任命的临床部分，从而允许一段时间的集中研究，以促进他的科学发展并扩大他做出重大研究贡献的潜力。 候选人处于一个高度支持的环境中，拥有优秀的导师和众多的智力发展机会。他是新成立的“炎症生物学”中心的教员。候选人的实验室位于中心及其核心实验室内，其中包括20，000平方英尺。新装修的最先进的开放式实验室设计设施。该中心的研究环境提供了一个动态的临界质量的研究人员提供了一个协同的环境，为候选人的研究。 核心研究项目的总体目标（一个为期五年的R 01资助8/1/02）是了解气道上皮细胞中iNOS降解的机制。本研究的主要目的是：1）鉴定人iNOS的降解途径。2)泛素化在诱导型一氧化氮合酶降解中可能作用的表征。3)阐明人iNOS的亚细胞定位。4)通过别构抑制剂调节iNOS降解的分析。候选人通过合作研究解决了三个额外的具体目标：I）确定iNOS泛素化的特定位点。II）阐明磷酸化在iNOS泛素化中的作用。III）鉴定负责iNOS泛素化的特异性泛素连接酶（E3）。将在上皮细胞系和原代气道支气管上皮细胞中进行研究。 提出的研究的理由是，一旦了解了iNOS的降解机制，就可以设计治疗策略来改变这些途径并调节iNOS的降解。这些研究的结果将增加我们对iNOS调节的细胞过程的理解，从而为未来旨在控制气道炎症疾病（如哮喘）中NO合成的研究奠定基础。

DESCRIPTION (provided by applicant): The candidate is a recently independent physician-scientist who has shown strong commitment to biomedical research. His long-term career goal is to develop a strong research program in molecular mechanisms of airway inflammation. His immediate research goals are to understand the molecular mechanisms of regulation NO synthesis by inducible nitric oxide synthase (iNOS) in airway cells and devise methods to control it. A successful R01 application has the candidate poised to continue his current studies, initiate new programs, acquire new skills and build new collaborations. The K02 award is needed to reduce the clinical component of the candidate's current appointment, thus allowing a period of intensive research focus to foster his scientific development and expand his potential to make significant research contributions. The candidate is in a highly supportive environment with excellent mentors and numerous opportunities for intellectual development. He is a faculty member of a newly established center for "Biology of Inflammation". The candidate's laboratory is within the center and its core laboratories, which comprise 20,000 sq, ft. of newly renovated state-of-the-art-open lab design facilities. The center's research environment offers a dynamic critical mass of investigators providing a synergistic environment for the candidate's research. The overall objective of the core research project (a five-year R01 funded 8/1/02) is to understand the mechanisms of iNOS degradation in airway epithelial cells. It consists of four specific aims: 1) Identification of human iNOS degradation pathway. 2) Characterization of possible role of ubiquitination in iNOS degradation. 3) Elucidation of the subcellular localization of human iNOS. 4) Analysis of modulation of iNOS degradation by allosteric inhibitors. The candidate is addressing three additional specific aims through collaborative studies: I) Determination of specific sites of iNOS ubiquitination. II) Elucidation of Role of phosphorylation in iNOS ubiquitination. III) Identification of specific ubiquitin ligase (E3) responsible for iNOS ubiquitination. Studies will be conducted in epithelial cell lines and in primary airway bronchial epithelial cells. The rational for the proposed studies is that once the degradation mechanisms of iNOS are understood, therapeutic strategies can be designed to alter these pathways and modulate iNOS degradation. The results of these studies will increase our understanding of the cellular process of iNOS regulation and thus lay the groundwork for future studies aiming at controlling NO synthesis in diseases of airway inflammation such as asthma.