Structures, Mechanisms, and Biogenesis of Amine Oxidases

说明(申请人提供)：含铜胺氧化酶和赖氨酸氧化酶在自然界中分布广泛，参与生物性伯胺的代谢、结缔组织的成熟和许多其他生理过程。人血管黏附蛋白(hVAP-1)是一种胺类氧化酶，其活性直接参与细胞黏附。哺乳动物体内存在多种类赖氨酰氧化酶(LOXL)蛋白，这些蛋白可以替代赖氨酰氧化酶，但也可能在多种细胞过程中发挥作用，包括分化、增殖和运动。最近发现的生物胺和几种药物化合物可能被胺氧化酶代谢或抑制。提出了一种强调光谱学、动力学、定点突变和结晶学的多学科方法，以阐明定义胺氧化酶和赖氨酸氧化酶关键结构-功能关系的化学和生物学原理。一个主要的目标是确定关键的胺氧化酶的结构，包括赖氨酰氧化酶，以及胺氧化酶和赖氨酸氧化酶与抑制剂和底物类似物的复合体的结构。其他主要目标是确定胺氧化酶的活性部位结构、催化机制以及底物专一性和选择性抑制的分子基础，包括hVAP-1和LOXL。还提出了旨在解决TPQ和LTQ生物发生中悬而未决的机制问题的实验。此外，还将研究一种相关酶--半乳糖氧化酶的结构和生物发生。结合结构和机理数据将使人们能够详细了解这些待开发的重要酶的结构和功能。

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.