HOST GUEST METALLOPROTEIN MIMICS

宿主客体金属蛋白模拟物

• 批准号: 2022716
• 负责人: JAMES W CANARY
• 金额: $ 10.78万
• 依托单位: NEW YORK UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-12-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2022716
• 关键词: X ray crystallography; active sites; calorimetry; carbonate dehydratase; carboxylate; carboxypeptidase; chemical kinetics; enzyme inhibitors; enzyme substrate complex; hydrogen bond; potentiometry; sulfonamides; thermodynamics; water; zinc

The ultimate goal of this research project is the development of therapeutic agents for diseases that can be treated by inhibiting zinc hydrolytic enzymes, including rheumatoid and osteoarthritis, periodontitis, glaucoma, and cancer. The design of these agents requires a knowledge of (1) the mechanism of the enzyme and (2) the energies of binding interactions between the enzyme and an inhibitor. The catalytic mechanisms and inhibitor binding energies are the sum of many varied forces. It is difficult to measure the contribution of an individual type of interaction in an enzyme due to its size and complexity. This proposal focuses on two of the most highly studied zinc hydrolytic enzymes, carbonic anhydrase and carboxypeptidase A. It is believed that many other zinc enzymes are similar in structure and utilize similar mechanisms. We have chosen to address specifically the means by which enzymes activate water that is coordinated to zinc in the active site. Studies of carbonic anhydrase suggest that hydrophobic interactions, hydrogen bonds, and the presence of carboxylates near the zinc ion or the water affect enzyme activity and inhibitor binding, but the magnitude and the detailed physical basis for the effects are unknown. This proposal seeks to quantify the effects and establish how and why the stated interactions operate. In carboxypeptidase A, the overall mechanistic path is still a matter of controversy, to say nothing of the details of the path. There is presently no ligand that forms a complex with zinc and has ligands and geometry which closely resemble those found in CPA. This proposal contains a plan to develop such a ligand. Binding studies will be used to determine the energy of inhibitor coordination to zinc in the absence of secondary interactions in the enzyme active site. Once this goal has been achieved, ligands will be prepared which quantify the contribution of some of the known active site secondary interactions. Proposed mechanisms of action of carboxypeptidase A will be tested in carefully designed model complexes. The tools used to achieve the goals of this study include molecular design and synthesis of model coordination complexes. The complexes will be characterized by x-ray crystallography, potentiometry, calorimetry, kinetic studies, and other techniques.

该研究项目的最终目标是开发 可通过抑制锌来治疗的疾病的治疗剂 水解酶，包括类风湿和骨关节炎， 牙周炎、青光眼和癌症。这些代理的设计需要 了解 (1) 酶的机制和 (2) 酶的能量 酶和抑制剂之间的结合相互作用。催化 机制和抑制剂结合能是许多不同因素的总和 力量。很难衡量个体类型的贡献 由于其大小和复杂性，酶中的相互作用。这个提议 专注于两种研究最深入的锌水解酶， 碳酸酐酶和羧肽酶 A。据信还有许多其他酶 锌酶在结构上相似并且利用相似的机制。 我们选择专门解决酶激活的方式 与活性位点中的锌配位的水。碳酸的研究 脱水酶表明疏水相互作用、氢键和 锌离子或水附近存在的羧酸盐会影响酶 活性和抑制剂结合，但幅度和详细 影响的物理基础尚不清楚。该提案旨在 量化影响并确定所述相互作用的方式和原因 操作。在羧肽酶 A 中，总体机制路径仍然是 争议的问题，更不用说路径的细节了。有 目前还没有配体与锌形成络合物并且具有配体和 几何形状与 CPA 中的几何形状非常相似。该提案包含 开发这种配体的计划。结合研究将用于确定 在没有次级的情况下抑制剂与锌配位的能量 酶活性位点的相互作用。一旦这个目标实现了， 将制备配体，量化一些配体的贡献 已知的活性位点二次相互作用。拟议的作用机制 羧肽酶 A 将在精心设计的模型复合物中进行测试。 用于实现本研究目标的工具包括分子设计 和模型配位配合物的合成。这些综合体将是 通过 X 射线晶体学、电位测定法、量热测定法进行表征， 动力学研究和其他技术。