Cofactor-Dependent Amine Oxidations

辅因子依赖性胺氧化

基本信息

批准号：
7417522
负责人：
IRENE LEE
金额：
$ 18.88万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-07-01 至 2010-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7417522
关键词：
6-hydroxydopa quinone Accounting Active Sites Adhesions Aldehydes Alkylation Amines Arthritis Atherosclerosis Behavior Therapy Binding Binding Sites Biogenesis Biogenic Amines Biological Assay Biological Process Blood Vessels Bos taurus Cardiovascular Diseases Cations Cattle Cell division Chemicals Chemistry Class Collaborations Complex Complications of Diabetes Mellitus Copper Data Databases Deposition Detection Development Development, Other Diabetes Mellitus Diamines Digestion Disease Enzymes Family Fibrosis Fingerprint Flavins Future Genetic Grant Growth Human Immune System Diseases Individual Inflammation Inflammatory Investigation Kidney Kinetics Life Ligands Localized Mammals Mass Spectrum Analysis Mediating Membrane Metabolism Methods Modality Modeling Modification Molecular Monoamine Oxidase Multiple Sclerosis Nature Numbers Organism Oxidases Peptides Pharmacologic Substance Physiological Plants Plasma Play Polyamines Process Prodrugs Protein-Lysine 6-Oxidase Proteins Public Health Quinones Range Reaction Research Research Personnel Roentgen Rays Role Semicarbazides Sorting - Cell Movement Spectrometry Staging Structure Study models System Time Tissues Tyrosine Work amine oxidase analog base benzoquinone biological adaptation to stress carbamylhydrazine cofactor concept cytotoxic design diamino oxhydrase drug development human AOC3 protein immunoregulation inhibitor/antagonist interest molecular modeling novel novel strategies oxidation polyamine oxidase preference programs propargylamine protein structure stable isotope therapeutic target tool transamination

项目摘要

DESCRIPTION (provided by applicant): The copper-containing amine oxidases (CAOs) comprise a ubiquitous class of enzymes in all living organisms. In plants and mammals, CAOs play important roles in metabolizing biogenic amines involved in growth, cell division, differentiation, and the stress response, where metabolism in some cases has cytotoxic consequences. All CAOs utilize a quinone cofactor derived postranslationally from an active-site tyrosine residue to mediate a transaminative conversion of primary amines to aldehydes. Studies using cofactor models in the past grant periods have inspired the discovery of new types of enzyme-activated mechanisms of inhibition. These studies, along with development of other traditional inactivation strategies, have resulted in highly promising degrees of inhibitor selectivity. The selectivity being achieved is discriminating not only between the flavin- and quinone-dependent enzymes, but also among a range of CAOs that have different substrate-structure preferences. Since the time of the last renewal, the genetic basis of the two general classes of human CAOs has been revealed: (i) the trihydroxyphenylalanine quinone (TPQ)-dependent class AOC 1 (the human kidney diamine oxidase), AOC 2, and AOC 3, the latter coincidental with the human vascular adhesion protein-1 (HVAP-1), and (ii) four lysyl oxidase-like (LOXL) proteins in addition to classical LOX. In the next grant period, there will be increased focus on refining inhibitor selectivity for the human enzymes, particularly the kidney/placental diamine oxidase, the soluble and tissue-bound semicarbazide- sensitive monoamine oxidases, and lysyl oxidase. The new aims include the development of sensitive fluorometric assays for detecting and localizing mammalian CAOs, an exploration of new inhibitor selectivity and prodrug constructs, the use of x-ray crystallographic information (through collaboration) for refinement of substrate and inhibitor binding models constructed by molecular modeling, and the use of spectroscopy and mass spectrometry to identify the structural basis of irreversible inhibitor action. Studies on the nature of biogenesis of the (TPQ) cofactor will additionally be completed. RELEVANCE OF THIS RESEARCH TO PUBLIC HEALTH The copper amine oxidases have become important pharmaceutical targets on account of the emerging recognition of their key physiological roles in aspects of inflammation, immune modulation, fibrosis in arthritis and atherosclerosis, and in late-stage complications of diabetes and cardiovascular disease. It is thus important to understand the functioning of these enzymes, both from a mechanistic view and in terms of the structural basis of substrate recognition and the action of inhibitors that can serve a leads in eventual drug development.

描述（由申请人提供）：含铜胺氧化酶（CAO）包含所有生物体中普遍存在的一类酶。在植物和哺乳动物中，CAO 在参与生长、细胞分裂、分化和应激反应的生物胺代谢中发挥着重要作用，其中代谢在某些情况下具有细胞毒性后果。所有 CAO 都利用翻译后衍生自活性位点酪氨酸残基的醌辅因子来介导伯胺向醛的转氨基转化。在过去的资助期间使用辅因子模型的研究启发了新型酶激活抑制机制的发现。这些研究以及其他传统灭活策略的发展已经产生了非常有希望的抑制剂选择性程度。所实现的选择性不仅可以区分黄素依赖性酶和醌依赖性酶，还可以区分具有不同底物结构偏好的一系列 CAO。自上次更新以来，人类 CAO 的两个一般类别的遗传基础已被揭示：(i) 三羟基苯丙氨酸醌 (TPQ) 依赖性类别 AOC 1（人肾二胺氧化酶）、AOC 2 和 AOC 3，后者与人血管粘附蛋白-1（HVAP-1）一致，以及（ii）除了经典的赖氨酰氧化酶样（LOXL）蛋白之外液氧。在下一个资助期内，将更加关注改进人类酶的抑制剂选择性，特别是肾脏/胎盘二胺氧化酶、可溶性和组织结合的氨基脲敏感单胺氧化酶和赖氨酰氧化酶。新的目标包括开发用于检测和定位哺乳动物 CAO 的灵敏荧光测定法、探索新的抑制剂选择性和前药结构、使用 X 射线晶体学信息（通过合作）来细化由分子构建的底物和抑制剂结合模型。建模，并使用光谱和质谱来确定不可逆抑制剂作用的结构基础。此外，还将完成对 (TPQ) 辅因子生物发生性质的研究。本研究与公众健康的相关性由于人们逐渐认识到铜胺氧化酶在炎症、免疫调节、关节炎和动脉粥样硬化纤维化以及糖尿病和心血管疾病晚期并发症方面的关键生理作用，铜胺氧化酶已成为重要的药物靶点。因此，从机制角度和底物识别的结构基础以及可以在最终药物开发中发挥作用的抑制剂的作用方面了解这些酶的功能非常重要。