Structures, Mechanisms, and Biogenesis of Amine Oxidases

胺氧化酶的结构、机制和生物发生

• 批准号: 7196174
• 负责人: DAVID M. DOOLEY
• 金额: $ 33.5万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7196174
• 关键词: Drosophilidae; X ray crystallography; amine oxidase (copper); amine oxidoreductase; chemical kinetics; cofactor; copper; enzyme biosynthesis; enzyme induction /repression; enzyme inhibitors; enzyme mechanism; enzyme substrate analog; enzyme substrate complex; galactose oxidase; intermolecular interaction; molecular site; protein purification; protein structure function; site directed mutagenesis; spectrometry; tyrosine

DESCRIPTION (provided by applicant): Copper-containing amine oxidases and lysyl oxidases are widely distributed in nature and are involved in the metabolism of biogenic primary amines, in the maturation of connective tissue, and numerous other physiological processes. The human vascular adhesion protein (hVAP-1) is an amine oxidase, and its oxidase activity is directly involved in cellular adhesion. Multiple "lysyl oxidase like" (LOXL) proteins have been recognized in mammals, and these proteins may substitute for lysyl oxidase, but may also have roles in multiple cellular processes, including differentiation, proliferation, and motility. It appears that recently recognized biogenic amines and several pharmaceutical compounds may be metabolized by or inhibit amine oxidases. A multidisciplinary approach emphasizing spectroscopy, kinetics, site directed mutagenesis, and crystallography is proposed to elucidate the chemical and biological principles that define key structure- function relationships in amine oxidases and lysyl oxidases. A major goal is to determine the structures of key amine oxidases, including a lysyl oxidase, and of amine oxidase and lysyl oxidase complexes with inhibitors and substrate analogues. Other major goals are to define the active-site structures, catalytic mechanisms, and the molecular bases for substrate specificity and selective inhibition in amine oxidases, including hVAP-1 and LOXLs. Experiments designed to resolve outstanding mechanistic questions in the biogenesis of TPQ and LTQ are also proposed. In addition, the structure and biogenesis of a related enzyme, galactose oxidase, will be examined. Combining structural and mechanistic data will permit a detailed understanding of the structure and function of these important enzymes to be developed.

描述（由申请人提供）：含铜的胺氧化酶和赖氨酸氧化酶在自然界中广泛分布，并参与了生物原代胺的代谢，结缔组织的成熟以及许多其他生理过程。人血管粘附蛋白（HVAP-1）是一种胺氧化酶，其氧化酶活性直接参与细胞粘附。在哺乳动物中已经识别出多种“赖氨酸氧化酶”（LOXL）蛋白，这些蛋白可能代替赖氨酸氧化酶，但也可能在多种细胞过程中起作用，包括分化，增殖和运动。看来最近识别的生物胺和几种药物化合物可以通过或抑制胺氧化酶代谢。提出了一种多学科方法，该方法强调光谱，动力学，定位诱变和晶体学以阐明定义胺氧化酶和赖氨酸氧化酶中关键结构关系的化学和生物学原理。一个主要目标是确定关键胺氧化酶的结构，包括赖氨酸氧化酶，以及与抑制剂和底物类似物的胺氧化酶和赖氨酸氧化酶复合物的结构。其他主要目标是定义活性位点结构，催化机制以及胺氧化酶（包括HVAP-1和LOXL）的底物特异性和选择性抑制的分子碱基。还提出了旨在解决TPQ和LTQ生物发生中出色的机械问题的实验。另外，将研究相关酶的结构和生物发生。结合结构和机械数据将允许对这些重要酶的结构和功能有详细的了解。