SPECTROSCOPIC STUDIES OF ENZYME-SUBSTRATE COMPLEXES

酶-底物复合物的光谱研究

• 批准号: 3288903
• 负责人: GEORGE H REED
• 金额: $ 16.27万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-01-01 至 1990-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3288903
• 关键词: adenosine triphosphate; cations; electron spin resonance spectroscopy; enzyme induction /repression; enzyme structure; enzyme substrate; enzyme substrate analog; enzyme substrate complex; infrared spectrometry; magnesium; manganese; molecular site; nuclear magnetic resonance spectroscopy; phosphoglycerate kinase; pyruvate kinase; stereochemistry; vanadium

Enzymes that catalyze the synthesis, utilization, and interconversion of phosphorylated-metabolites usually require specific activation by inorganic cations, and reactions involving metal nucleotide substrates or effectors are of central importance in a wide varity of cellular processes. Understanding the roles of metal ions in the respective catalytic activations, and in the overall mechanisms for these reactions requires detailed information on the binding interactions that occur between the metal ion, substrates, and protein in the reactive complexes. The goals of this project are to determine the coordination schemes in enzyme-metal ion-substrate and inhibitor complexes in order to understand the relationships between specific binding interactions and chemical activation of these molecules and the strategies that are exploited in the catalytic mechanisms. The proposed research will use electron paramagnetic resonance, nuclear magnetic resonance, and vibrational spectroscopy, together with isotopically labeled molecules (substrates and analogue), to determine the binding schemes between metal ion, substrate and protein at the active sites of several enzymes that catalyze phosphotransfer reactions. The experiments will also provide information on the stereospecificities of enzyme active sites and on the mode of action of specific inhibitory compounds. Knowledge of the structures of enzyme-substrate complexes is essential to the rational design of specific inhibitors that can be used as therapeutic agents, or as biochemical and physiological reagents to control, selectively, reactions or metabolic pathways. The information is also of fundamental importance in understanding the mechanisms for biological catalysis.

催化合成，利用和相互转换的酶 磷酸化的量代谢物通常需要通过无机激活 阳离子以及涉及金属核苷酸底物或效应子的反应 在各种细胞过程中至关重要。 了解金属离子在相应催化中的作用 激活以及这些反应的总体机制需要 有关绑定相互作用的详细信息 反应性复合物中的金属离子，底物和蛋白质。 目标 该项目旨在确定酶 - 金属中的协调方案 离子 - 基底和抑制剂复合物，以了解 特定结合相互作用与化学激活之间的关系 这些分子和在催化中利用的策略 机制。 拟议的研究将使用电子顺磁性 共振，核磁共振和振动光谱， 与同位素标记的分子（底物和类似物）一起 确定金属离子，底物和蛋白质之间的结合方案 几种催化磷酸转移的酶的活性位点 反应。 实验还将提供有关 酶有源位点的立体特异性以及在作用方式上 特定的抑制化合物。 了解结构 酶 - 基底络合物对于特定的理性设计至关重要 可以用作治疗剂的抑制剂，也可以用作生化和 有选择地控制反应或代谢的生理试剂 途径。 该信息在 了解生物催化的机制。