METALLOENZYME DYNAMICS AND MECHANISM

我们研究的目的是研究反应动力学 催化血红素酶，并获得有关结构的信息， 在催化过程中形成的催化中间体。 获得的信息 将用于制定具体的化学作用机制， 内切酶 这些研究将集中在辣根过氧化物酶，过氧化氢酶， 细胞色素C过氧化物酶和乳过氧化物酶。 还将进行比较 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 具有本质意义 最重要的是活性中心结构的 酶在这些中间体，因为这揭示了催化是如何带来的 关于. 为了实现这些目标，目前正在将两种方法结合起来。 停流冷冻酶学被用于检测和稳定特定的 溶液中的中间体。 采用共振拉曼光谱 选择性地检查血红素基团的振动光谱， 催化中间体 共振拉曼光谱提供了 关于血红素铁的氧化和自旋状态，性质和 铁轴配体键的强度和电子分布 血红素基团和轴向配体。 根据这些数据，详细的机制 将制定这些过氧化物酶的行动。

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.