Coenzyme A Analogs as Probes of Enzymatic Reactions

辅酶 A 类似物作为酶反应的探针

• 批准号: 9321371
• 负责人: Dale Drueckhammer
• 金额: $ 30万
• 依托单位: Stanford University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-15 至 1997-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9321371&HistoricalAwards=false
• 关键词: Coenzyme; Analogs; Probes; Enzymatic; Reactions

9321371 Drueckhammer This project will use synthetic Coenzyme A (CoA) analogs to study fundamental mechanistic questions in enzyme catalyzed reactions of CoA esters. This work is based on methodology developed in the P.I.'s lab for the convenient synthesis of CoA/CoA ester analogs. The largest effort will be directed at probing the involvement of enolate and/or enol intermediates in enzyme catalyzed reactions of acetyl-CoA and other CoA esters. Stable analogs of the enolate and enol forms of acetyl-CoA will be prepared, and studied as inhibitors of citrate synthase. The protonation state of these analogs in isotopically enriched form bound to the enzyme will be studied by NMR and in some cases FTIR. Similar studies will be conducted with related enzymes, including malate synthase, crotonase, and acetyl-CoA dependent acetyltransferases. This project is expected to provide new insights into the enzyme catalyzed reactions of CoA esters and general principles of enzyme reaction mechanisms. %%% Dr. Drueckhammer will study the reactions of enzymes which require Coenzyme A for their function. Enzymes are catalysts which carry out chemical reactions in biological systems. Understanding how these enzymes function is at the heart of understanding biological processes at a fundamental chemical level. About 4% of all enzymes require coenzyme A (CoA) for their catalytic activity. CoA serves as a reactant or a reactant carrier in these enzymatic reactions. CoA is also synthesized in biological systems by a series of enzyme catalyzed reactions. In previous work in this lab, a method was developed for using the enzymes which make CoA to make altered forms (analogs) of natural CoA. This research will use this method to make a series of CoA analogs designed to serve as probes for studying the reactions of CoA utilizing enzymes. This will help reveal how enzymes form chemical bonds to make complex biological molecules from simple building blocks, and how enzymes break down com plex compounds in food to produce biological energy. ***

9321371德鲁克哈默这个项目将使用合成辅酶A(CoA)类似物来研究辅酶A酯的酶催化反应中的基本机理问题。这项工作是基于P.I.S实验室为方便合成辅酶A/辅酶A酯类似物而开发的方法学。最大的努力将集中在探索烯醇和/或烯醇中间体在乙酰辅酶A和其他辅酶A酯的酶催化反应中的参与。将制备稳定的烯醇和烯醇形式的乙酰辅酶A类似物，并将其作为柠檬酸合成酶的抑制剂进行研究。核磁共振和某些情况下的FTIR将研究这些以同位素富集态结合到酶上的类似物的质子化状态。将对相关酶进行类似的研究，包括苹果酸合成酶、巴豆酶和乙酰辅酶A依赖的乙酰转移酶。该项目有望对辅酶A酯的酶催化反应和酶反应机理的一般原理提供新的见解。德鲁克哈默博士将研究需要辅酶A才能发挥作用的酶的反应。酶是在生物系统中进行化学反应的催化剂。了解这些酶的功能是在基础化学水平上了解生物过程的核心。大约4%的酶需要辅酶A(CoA)才能发挥催化活性。辅酶A在这些酶促反应中作为反应物或反应物载体。辅酶A也是在生物体系中通过一系列酶催化反应合成的。在本实验室以前的工作中，开发了一种方法，用于使用产生辅酶A的酶来生成天然辅酶A的改变形式(类似物)。本研究将利用这种方法制作一系列辅酶A类似物，作为研究辅酶A利用酶的反应的探针。这将有助于揭示酶是如何从简单的积木形成化学键来制造复杂的生物分子，以及酶是如何分解食物中的复杂化合物来产生生物能量的。***