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: 6893671
• 负责人: RAEGAN D HUNT
• 金额: $ 2.63万
• 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 2006-10-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6893671
• 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）是由三种结构上不同的一氧化氮合酶（NOS）从L-精氨酸和分子氧合成的气态信使：神经元（NNOS），内皮（ENOS）（ENOS）和诱导（Inos）。在正常情况下，没有在体内平衡中扮演关键角色。在中风中，NNOS没有过多生产会导致明显的神经毒性，而ENOS产生的没有有益于通过血管舒张和抑制血小板聚集的恢复血流，但NNOS是中风疗法的一个极好的分子靶标，但必须明确地选择抑制剂，因此不能以nnos isof的形式被构成阳性，从而构成了阳性构成。该研究项目的基本假设是1）NNOS和ENOS同工型的比较结构研究将识别差异，这可以针对同工型特异性抑制NNOS和2）对抑制天然NOS抑制剂抑制的结构基础的理解可以提供新的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测三个具体目的：1）确定NNOS催化血红素结构域的三维结构，2）NNOS的表征和比较NNOS特定的同工式衍生物结合了NNOS和ENOS Home域与NNOS和ENOS Homeain rout toins nnos和Enos Homeain的概念的确定。这些研究将为NNOS同工型特定药物设计提供基本科学框架。