EPR AND RESONANCE RAMAN SPECTROSCOPY

在这个核心中描述的光谱研究的长期目标 项目是使用从X射线获得的结构信息 前列腺素H合成酶和细胞色素的结晶学研究 C氧化酶(CcOX)作为探索其催化细节的平台 机制，并开发新的光谱方法来研究 生物系统中的金属中心。振动光谱学，共振 拉曼光谱和傅里叶变换红外光谱，它们提供了有关键的详细信息 强度，将扩展到金属-配体键合的能量区域 当与小体积、停流相结合时，可以更好地表征 动力学技术，这些光学方法提供了一条独特的途径 表征催化中间体。先进的EPR方法将 在这项核心工程中使用，允许一种测量波形功能 它包含一个未配对的电子自旋，并获得了对 催化位的电子结构。电子病历开发工作 将在这一核心项目中承担的工作包括开发 提高我们能力的脉冲-微波/脉冲磁场方法 为了解决高自旋顺磁金属中心的超精细耦合， 几种新的相干转移脉冲EPR方法的应用 蛋白质自由基构象和配基超精细的测定 联结，以及并行发展的分析战略，以支持 这些测量值。因为这篇文章中描述的开发工作 提案是由针对CcOX和 PGHS，在此项目过程中定义的方法学将是 通常对生物医学界有用。

The long-range goal of the spectroscopic studies described in this core project are to use the structural information gained from X-ray crystallographic studies of Prostaglandin H Synthase (PGHS) and Cytochrome c Oxidase (CcOX) as platforms for exploring the details of their catalytic mechanisms, and to develop new spectroscopic methods for the study of metal centers in biological systems. Vibrational spectroscopies, Resonance Raman and FTIR, which provide detailed information regarding bond strengths, will be extended to energy regimes where metal-ligand bonding can be better characterized When combined with small-volume, stop-flow kinetic techniques, these optical methods provide a unique pathway for characterizing catalytic intermediates. The advanced EPR methods that will be utilized in this core project, allow one to measure to wave function that house an unpaired electron spin and gain an understanding of the electronic structure of a catalytic site. The EPR development work that will be undertaken in this core project includes the development of a pulse-microwave/pulse-magnetic field method for improving our capability to resolve hyperfine couplings for high-spin paramagnetic metal centers, the application of several new coherence transfer pulsed EPR methods for the measurement of protein radical conformation and ligand hyperfine couplings, and the parallel development of analysis strategies to support these measurements. Because the development work described in this proposal is driven by specific questions being addressed for CcOX and PGHS, the methodologies defined during the course of this project will be generally useful to the biomedical community.