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.

这个奖项由无机，生物无机和有机化学和分子生物化学计划支持理查德教授B的研究。Holz在犹他州州立大学的研究中，确定了来自解蛋白气单胞菌的亮氨酸氨肽酶（AAP）和来自嗜热单胞菌的dapE编码的N-琥珀酰-L，L-二氨基庚二酸脱琥珀酰酶（DapE）的详细催化机制。并设计和合成有效的DapE抑制剂。 .这项研究涉及一个跨学科的方法，结合生物化学，光谱和X射线晶体学方法和DapE的小分子抑制剂的合成。 本工作涉及五个关键领域：（1）AAP活性中心的哪些残基起催化作用？(2)Laue X射线衍射方法在AAP催化机理研究中的应用。(3)底物和过渡态类似物抑制剂如何与DapE相互作用的表征。(4)DapE中提议的活性位点残基的改变的酶的制备。(5)AAP和DapE水解酶含有双核金属中心的详细作用机制的发展是许多生物过程的核心，因此，它们的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发挥重要作用。 几种双核氨肽酶具有广泛的底物特异性，广泛分布于动植物组织中。 由于它们在蛋白质和生物活性肽（包括激素）的降解中的作用，它们的生物学和医学意义是广泛的。了解亮氨酸氨肽酶的反应机制的重要性是由以下事实强调的：亮氨酸氨肽酶活性已在肿瘤细胞表面上观察到，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通过抑制亮氨酸氨肽酶活性显著降低男性HIV感染。 对于DapE，对所有目前可用的抗生素具有抗性的几种病原性细菌菌株的出现强调了它们的重要性，因为mDAP/赖氨酸生物合成途径的两种产物mDAP和赖氨酸是革兰氏阴性和大多数革兰氏阳性细菌的肽聚糖细胞壁的必需组分。这项研究旨在详细了解双核金属中心催化的水解反应，这可能会导致新抗生素的开发，并支持本科生，研究生和博士后的发展。