NEAR ULTRAVIOLET RAMAN STUDIES OF CYTOCHROME P450

细胞色素 P450 的近紫外拉曼研究

• 批准号: 3233335
• 负责人: Paul M. Champion
• 金额: $ 15.66万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-06-01 至 1993-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3233335
• 关键词: Pseudomonas; Raman spectrometry; acidity /alkalinity; bacterial proteins; chemical structure function; cytochrome P450; cytochrome c; electron spin resonance spectroscopy; hemoglobin; hemoprotein; ion transport; laser spectrometry; ligands; mutagens; myoglobin; photolysis; ultraviolet spectrometry

We have observed detailed and informative resonance Raman spectra from heme proteins such as cytochrome P450, cytochrome c, myoglobin and hemoglobin. Laser excitation in the deep blue and near ultraviolet resonant absorption bands leads to greatly enhanced signal intensities associated with the heme prosthetic group. We aim to extend our understanding of the structure and function of cytochrome P450 and other heme protein systems by the use of Raman and other rapidly developing laser based spectroscopies. These studies will employ Raman excitation profile measurements that allow key electron-nuclear coupling parameters to be extracted from the combined absorption and Raman data. Careful studies of the absorption and Raman lineshapes will also be carried out as a function of temperature. Sample perturbations involving site directed mutagenesis, pH and optical pumping are also scheduled. Time resolved protein dynamics will be studied in both the nanosecond and picosecond regimes. Ligand photolysis and electron transport phenomena along with thermal relaxation and non-radiative decay processes will be investigated. Single crystal samples of myoglobin and cytochrome P450cam will also be studied as a function of orientation and excitation wavelength. Various ligands that induce z-polarized charge transfer excitations will be complexed to the crystals in order to examine the details of the polarizability tensor. Theoretical work involving ligand binding to heme proteins, electron transfer and resonant light- matter interactions will also be pursued. Collaborative experiments involving microwave conductivity, electron spin resonance and deep ultraviolet Raman scattering will be initiated. This overall project has wide range of health related implications involving cytochrome P450 in particular and heme proteins in general. Many chemotherapeutic agents as well as polycyclic carcinogens are metabolized by P450 systems. Moreover, all metabolic disorders involving a P450 protein must relate to this research. Certainly a fundamental understanding of the active site structure and function of P450 at the molecular level will result in a better insight for those concerned with treatment of these disorders at any level.

我们观察到了血红素的详细且信息丰富的共振拉曼光谱 蛋白质，例如细胞色素 P450、细胞色素 c、肌红蛋白和血红蛋白。 激光激发深蓝色和近紫外共振吸收 带导致与血红素相关的信号强度大大增强 假肢组。 我们的目标是扩展我们对结构和 细胞色素 P450 和其他血红素蛋白系统的功能 拉曼和其他快速发展的基于激光的光谱学。 这些 研究将采用拉曼激发剖面测量，允许关键 从组合中提取电子-核耦合参数 吸收和拉曼数据。 仔细研究吸收和拉曼 线形状也将作为温度的函数进行。 样本 涉及定点诱变、pH 和光泵浦的扰动 也已安排。 时间分辨蛋白质动力学将在以下两个方面进行研究 纳秒和皮秒状态。 配体光解和电子 传输现象以及热弛豫和非辐射衰变 过程将受到调查。 肌红蛋白的单晶样品和 细胞色素 P450cam 也将作为方向和功能的函数进行研究 激发波长。 诱导 z 极化电荷的各种配体 转移激发将与晶体复合以检查 极化张量的详细信息。 理论工作涉及 配体与血红素蛋白的结合、电子转移和共振光- 物质相互作用也将被追求。 合作实验 涉及微波传导、电子自旋共振和深 将启动紫外拉曼散射。 本次总体工程有 涉及细胞色素 P450 的广泛健康相关影响 特别是血红素蛋白。 许多化疗 药物以及多环致癌物均由 P450 系统代谢。 此外，所有涉及 P450 蛋白的代谢紊乱都必须与 这项研究。 当然是对活性位点的基本了解 P450 在分子水平上的结构和功能将导致 为那些关心这些疾病治疗的人提供更好的见解 等级。