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REGULATION OF MICROBIAL OXIDATION REACTIONS

微生物氧化反应的调节

基本信息

批准号：
3269482
负责人：
JULIAN A PETERSON
金额：
$ 14.65万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
1975
资助国家：
美国
起止时间：
1975-05-01 至 1991-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3269482
关键词：
Pseudomonas bacterial proteins computer simulation cytochrome P450 electron spin resonance spectroscopy electron transport enzyme mechanism enzyme substrate complex ionic strengths laboratory rabbit laboratory rat oxidation reduction reaction radiotracer redoxin respiratory oxygenation stereochemistry stop flow technique temperature jump

项目摘要

The long term goal of this research program is to attain an understanding of the mechanism and regulation of cytochrome P-450 catalyzed monooxygenation reactions. These enzymes are important because they are involved in the oxidation of drugs, synthesis of steroid hormones, activation of carcinogens and metabolism of polyunsaturated fatty acids. The research has focused on the bacterial cytochrome P-450 which catalyzes the oxidation of camphor because this enzyme system can be readily isolated in pure form in quantities which are sufficient to be used for this study. In the coming grant period the research will be focused on the identification of the transient intermediates which are formed during the reduction of cytochrome P-450 by reduced putidaredoxin, the physiological reductant. The kinetics of formation and decay of these intermediates will be measured with stopped-flow and temperature jump spectrophotometry. The intermediates will be trapped by freeze-quench techniques so that the EPR spectrum can be examined and the identity of the intermediates confirmed. The kinetics and control of the overall reaction of cytochrome P-450 will be evaluated with an appropriate computer model so that the precise control of this important enzyme system can be better appreciated.

这项研究计划的长期目标是了解细胞色素P-450催化的机制和调控单氧化反应这些酶很重要，因为它们是参与药物的氧化，类固醇激素的合成，致癌物质的活化和多不饱和脂肪酸的代谢。这项研究集中在细菌细胞色素P-450上，樟脑的氧化，因为这种酶系统可以容易地分离以纯的形式，其量足以用于本研究。在即将到来的资助期内，研究将集中在鉴定过程中形成的瞬时中间体通过还原的putidaredoxin还原细胞色素P-450，还原剂这些中间体的形成和衰变的动力学将采用停流温度跃变分光光度法测定。的中间体将被冷冻淬灭技术捕获，可以检查光谱并确认中间体的身份。细胞色素P-450的总体反应的动力学和控制将使用适当的计算机模型进行评估，以便精确控制这个重要的酶系统可以更好地理解。