Mechanism Based Design of Novel Inhibitors of Dinuclear Hydrolases

基于机制的双核水解酶新型抑制剂的设计

• 批准号: 0240810
• 负责人: Richard Holz
• 金额: $ 34.5万
• 依托单位: Utah State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-01-15 至 2005-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0240810&HistoricalAwards=false
• 关键词: Mechanism; Based; Design; Novel; Inhibitors

This award by the Inorganic, Bioinorganic and Organometallic Chemistry and the Molecular Biochemistry programs supports research by Professor Richard B. Holz at Utah State University to determine a detailed catalytic mechanism for the leucine aminopeptidase from Aeromonas proteolytica (AAP) and the dapE-encoded N-succinyl-L,L-diaminopimelic acid desuccinylase (DapE) from H. influenzae and to design and syntheize potent inhibitors of DapE. . This research involves an interdisciplinary approach that incorporates biochemical, spectroscopic, and X-ray crystallographic methods and the synthesis of small molecule inhibitors of DapE. The work addresses five key areas: (1) Which residues in the active site of AAP play catalytically important roles? (2) The development of Laue X-ray diffraction methods to study the catalytic mechanism of AAP. (3) Characterization of how substrate- and transition-state analog inhibitors interact with DapE. (4) Preparation of altered enzymes of proposed active site residues in DapE. (5) The development of a detailed mechanism of action for AAP and DapEHydrolases that contain dinuclear metal centers are central to numerous biological processes and, consequently, characterization of their structure and function is a problem of outstanding importance. They play important roles in tissue repair, protein maturation, hormone level regulation, cell-cycle control, as well as cell wall synthesis. Several dinuclear aminopeptidases have broad substrate specificities and are widely distributed in both plant and animal tissues. Their biological and medicinal significance is extensive due to their roles in the degradation of proteins and biologically active peptides, including hormones. The importance of understanding the reaction mechanism of leucine aminopeptidases is underscored by the fact that leucine aminopeptidase activity has been observed on the surface of tumor cells, and such activity is known to play key roles in oncogenesis. In addition, the naturally occurring peptide analog inhibitor, bestatin, significantly decreases HIV infection in males by inhibiting leucine aminopeptidase activity. For DapE's, the emergence of several pathogenic bacterial strains that are resistant to all currently available antibiotics emphasize their importance, since both products of the mDAP/lysine biosynthetic pathway, mDAP and lysine, are essential components of the peptidoglycan cell wall for Gram-negative and most Gram-positive bacteria. This research aims toward a detailed understanding of hydrolytic reactions catalyzed by dinuclear metal centers, which may lead to the development of new antibiotics and also supports the development of undergraduate, graduate and postdoctoral students.

该奖项获得了无机，生物无机和有机金属化学和分子生物化学计划，支持犹他州立大学的Richard B. Holz教授的研究，以确定从气氛中的亮氨酸氨基肽酶的详细催化机制。 （Dape）来自H. h。threnzae，并设计和合成DAPE的有效抑制剂。 。这项研究涉及一种跨学科方法，该方法结合了生化，光谱和X射线晶体学方法以及DAPE小分子抑制剂的合成。 该作品解决了五个关键领域：（1）AAP活跃部位中哪些残留物在催化重要的角色？ （2）LAUE X射线衍射方法的发展以研究AAP的催化机制。 （3）表征底物和过渡态类似物抑制剂与DAPE相互作用的表征。 （4）制备DAPE中提出的活性位点残基的酶改变的酶。 （5）开发包含双核金属中心的AAP和Dapehydrolases的详细作用机理对于众多生物学过程至关重要，因此，其结构和功能的表征是一个非常重要的问题。 它们在组织修复，蛋白质成熟，激素水平调节，细胞周期控制以及细胞壁合成中起着重要作用。 几种双核氨基肽酶具有广泛的底物特异性，并且在动植物组织中分布广泛。 由于它们在蛋白质降解和包括激素在内的生物活性肽的降解中的作用，它们的生物学和药物意义是广泛的。理解亮氨酸氨基肽酶反应机制的重要性是由于已经在肿瘤细胞表面观察到了亮氨酸氨基肽酶活性的事实，并且已知这种活性在肿瘤发生中起关键作用。 此外，天然存在的肽类似物抑制剂Bestatin通过抑制亮氨酸氨基肽酶活性可显着降低雄性HIV感染。 对于Dape而言，由于MDAP/Lysine生物合成途径，MDAP和赖氨酸的两种产物都是肽聚糖细胞壁的基本成分，这些抗生素的几种病原细菌菌株的出现都强调了它们的重要性。这项研究旨在详细了解围栏金属中心催化的水解反应，这可能导致新的抗生素的发展，并支持本科，研究生和博士后学生的发展。