Studies on the mechanism of Pyridine Nucleotide Dependent Enzymes

吡啶核苷酸依赖性酶的作用机制研究

• 批准号: 8818529
• 负责人: Bruce Anderson
• 金额: $ 16.9万
• 依托单位: Virginia Polytechnic Institute and State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-02-01 至 1993-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8818529&HistoricalAwards=false
• 关键词: Studies; mechanism; Pyridine; Nucleotide; Dependent

The objectives here are to gain a better understanding of the mechanisms by which pyridine nucleotides function as coenzymes in dehydrogenase-catalyzed oxidation-reduction reactions and as substrates in NAD glycohydrolase-catalyzed hydrolyses and ADP- ribosylation reactions. These studies will include the isolation and purification of pyridine nucleotide dependent enzymes, the investigation of chemical, physical and kinetic properties of these enzymes and studies of the binding processes involved in the proper orientation of substrates and products. Structural analogs of the pyridine and nucleotides will be synthesized, chemically characterized, and applied in studies of selective interactions important to enzymic processes. Some of these derivatives will be designed as fluorescent probes while others will be designed to function as site-labeling reagents or suicide substrates. These studies will employ spectrophotometry, fluorimetry, titrimentry, liquid chromatography, and a number of general biochemical and chemical techniques. The overall importance of coenzyme analogs as experimental tools for he study of many biochemical processes is well-documented. The proposed new series of coenzyme analogs having unique properties will provide additional reagents applicable to biochemical studies. In addition to the proposed use of these compounds to elucidate enzymic mechanisms involved in NAD metabolism in H. influenzae, these analogs should find an even broader application in mechanistic studies of the many essential pyridine nucleotide-dependent enzymes.

这里的目标是更好地了解 吡啶核苷酸作为辅酶的作用机制 在脱氢酶催化的氧化还原反应中， 底物NAD糖水解酶催化水解和ADP- 核糖基化反应 这些研究将包括 吡啶核苷酸依赖性分离纯化 酶，化学、物理和动力学研究 这些酶的性质和结合的研究 涉及衬底的正确取向的过程， 产品. 吡啶和核苷酸的结构类似物 将被合成，化学表征，并应用于 选择性相互作用的研究对酶的 流程. 其中一些衍生品将被设计为 荧光探针，而其他探针将被设计为用作 位点标记试剂或自杀底物。 这些研究 将采用分光光度法，荧光法，滴定法，液体 色谱法，和一些一般的生物化学和 化学技术 辅酶类似物作为实验性药物的整体重要性 研究许多生物化学过程的工具是 记录良好 提出的新系列辅酶类似物 具有独特的性质将提供额外的试剂 适用于生物化学研究。 除了拟议的 这些化合物用于阐明涉及的酶机制的用途 在H.流感病毒，这些类似物应该找到一个 甚至在许多机械研究中的更广泛应用 必需的吡啶核苷酸依赖酶。