METALLOENZYME DYNAMICS AND MECHANISM

金属酶动力学和机制

• 批准号: 3274679
• 负责人: HAROLD E VAN WART
• 金额: $ 7.84万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-01-01 至 1989-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3274679
• 关键词: Raman spectrometry; catalyst; chemical structure function; chromophore; conformation; cryoscience; enzyme structure; enzyme substrate; metalloenzyme; solutions; solvents

The objective of our research is to study the dynamics of reactions catalyzed by heme enzymes and to obtain information about the structures of catalytic intermediates formed during catalysis. The information obtained will be used to formulate specific chemical mechanisms of action for these enzymes. The studies will focus on horseradish peroxidase, catalase, cytochrome c peroxidase and lactoperoxidase. Comparisons will also be made with myoglobin. To elucidate the complete mechanism of an enzyme, a detailed description of the elementary steps in the catalytic pathway is essential. Of central importance is the active site structure of the enzyme in these intermediates, since this reveals how catalysis is brought about. To achieve these goals, two current approaches are being combined. Stopped flow cryoenzymology is being used to detect and stabilize specific intermediates in solution. Resonance Raman spectroscopy is being employed to selectively examine the vibrational spectrum of the heme group in the catalytic intermediates. The resonance Raman spectra provide information about the oxidation and spin state of the heme iron, the nature and strength of the iron-axial ligand bonds and the electron distribution of the heme group and the axial ligands. From this data, detailed mechanisms of action will be formulated for these peroxidases.

我们研究的目的是研究反应的动力学 由血红素酶催化，并获取有关结构的信息 催化过程中形成的催化中间体。 获得的信息 将用于制定特定的作用化学机制 酶。 这些研究将重点放在辣根过氧化物酶，过氧化氢酶， 细胞色素c过氧化物酶和乳糖氧化酶。 还将进行比较 与肌红蛋白。 为了阐明酶的完整机制 催化途径中基本步骤的详细描述是 基本的。 至关重要的是主动地点结构 这些中间体中的酶，因为这揭示了如何带来催化 关于。 为了实现这些目标，正在合并两种当前的方法。 停止流量的冷冻酶学用于检测和稳定特定 溶液中的中间体。 共振拉曼光谱正在采用 有选择地检查血红素基团的振动光谱 催化中间体。 共振拉曼光谱提供信息 关于血红素铁的氧化和旋转状态，性质和 铁轴配体键的强度和电子分布 血红素组和轴向配体。 从这些数据，详细的机制 这些过氧化物酶将制定作用。