Structural Biology of Neuronal Nitric Oxide Synthase

神经元一氧化氮合酶的结构生物学

• 批准号: 7052802
• 负责人: RAEGAN D HUNT
• 金额: $ 0.72万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-15 至 2006-06-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7052802
• 关键词: X ray crystallography; active sites; affinity chromatography; caveolins; enzyme activity; enzyme inhibitors; enzyme structure; heme; isozymes; neurons; nitric oxide synthase; predoctoral investigator; protein protein interaction; protein purification

DESCRIPTION (provided by applicant): Nitric oxide (NO) is a gaseous messenger synthesized from L-arginine and molecular oxygen by three structurally distinct nitric oxide synthases (NOS): neuronal (nNOS), endothelial (eNOS), and inducible (iNOS). Under normal conditions, NO plays key roles in homeostasis. In stroke, NO overproduction by nNOS results in significant neurotoxicity, while NO generated by eNOS is beneficial to restoring blood flow via vasodilation and inhibition of platelet aggregation, nNOS is an excellent molecular target for stroke therapy, but inhibitors must be clearly selective for the nNOS isoform such that they do not block the positive effects resulting from NO production by eNOS. The hypotheses underlying this research project are that 1) comparative structural study of the nNOS and eNOS isoforms will identify differences, which can be targeted for isoform specific inhibition of nNOS and 2) an understanding of the structural basis of inhibition of a natural NOS inhibitor may provide novel NOS inhibition strategies. The structural basis of nNOS inhibition will be probed by high resolution x-ray crystallographic studies addressing three Specific Aims: 1) the determination of the three dimensional structure of the catalytic heme domain of nNOS, 2) characterization and comparison of nNOS specific isoform derivatives bound to the nNOS and eNOS home domain, and 3) determination of the NOS recognition site of a natural protein inhibitor, caveolin-1. These studies will provide the basic science framework for nNOS isoform specific drug design.

描述（由申请人提供）：一氧化氮 (NO) 是一种气体信使，由 L-精氨酸和分子氧通过三种结构不同的一氧化氮合酶 (NOS) 合成：神经元 (nNOS)、内皮 (eNOS) 和诱导型 (iNOS)。在正常情况下，NO 在体内平衡中发挥着关键作用。在中风中，nNOS 过量产生 NO 会导致显着的神经毒性，而 eNOS 产生的 NO 有利于通过血管舒张和抑制血小板聚集来恢复血流，nNOS 是中风治疗的绝佳分子靶点，但抑制剂必须对 nNOS 同工型具有明确的选择性，这样它们就不会阻碍 eNOS 产生 NO 所产生的积极作用。该研究项目的假设是：1）nNOS 和 eNOS 异构体的比较结构研究将识别差异，从而可以针对 nNOS 的异构体特异性抑制；2）了解天然 NOS 抑制剂抑制的结构基础可能会提供新的 NOS 抑制策略。 nNOS 抑制的结构基础将通过高分辨率 X 射线晶体学研究来探讨，解决三个具体目标：1）确定 nNOS 催化血红素结构域的三维结构，2）表征和比较与 nNOS 和 eNOS 主结构域结合的 nNOS 特异性亚型衍生物，以及 3）确定天然蛋白质抑制剂 Caveolin-1 的 NOS 识别位点。这些研究将为 nNOS 亚型特异性药物设计提供基础科学框架。