STRUCTURE OF CYTOCHROME P450 REDOX PARTNER COMPLEXES

细胞色素 P450 氧化还原伙伴复合物的结构

• 批准号: 6018443
• 负责人: Irina F Sevrioukova
• 金额: $ 3.68万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6018443
• 关键词: X ray crystallography; crystallization; cytochrome P450; enzyme complex; enzyme mechanism; enzyme structure; flavin mononucleotide; isozymes; oxidoreductase

Interaction of the P450 enzymes with their redox partners and the mechanism of electron transfer are the most important and intriguing problems in the area of P450 research. The crystal structures of four bacterial P450s, the catalytical domain of microsomal P450 reductase, and the NMR structure of putidaredoxin are known. However, knowledge of these structures does not directly answer where and how the proteins interact. Since interaction of other electron transfer proteins in crystallographic electron transfer complexes was shown to be specific and relevant to that occurring in a solution, we propose that crystallization of P450-redox partner complexes will enable the identification of interaction sites, conformation changes occurring upon binding, and the electron pathway from one co-factor to another. The P450cam electron transfer system and the heme/FMN-binding domain of flavocytochrome P450BM-3, a functional and structural analog to microsomal P450s and reductase, are the best candidates for crystallization trials of Class I and Class II monooxygenase systems, respectively. Having crystallized both the covalently linked heme/FMN-binding domain and the non-covalent complex between the separate heme-and FMN-binding domains of P450BM-3, we are in an excellent position to solve these structures, which will, undoubtedly, lead to our better understanding of the mechanism and function of microsomal P450-dependent monooxygenase systems.

P450酶与氧化还原伴侣的相互作用和 电子传递的机制是最重要，最吸引人的 P450研究领域的问题。四个的晶体结构 细菌P450，微粒体P450还原酶的催化结构域和 Putidaredoxin的NMR结构是已知的。但是，对这些知识 结构不能直接回答蛋白质的何处以及如何相互作用。 由于其他电子转移蛋白在晶体学中的相互作用 电子传输复合物被证明是特定的，并且与之相关 发生在溶液中，我们提出了P450-REDOX的结晶 合作伙伴建筑群将使交互作用位点的识别， 结合后发生的构象变化，电子途径从 一个共同因素是另一个。 P450CAM电子传输系统和 血红素/FMN结合结构域P450bm-3，功能性和功能性和 对微粒体P450和还原酶的结构类似物是最好的 I级和II类结晶试验的候选人 单加氧化酶系统分别。 两者都结晶了共价连接的血红素/FMN结合域和 单独的血红素和FMN结合域之间的非共价复合物 在P450BM-3中，我们在解决这些结构方面处于一个极好的位置 毫无疑问，这将导致我们对机制的更好理解 微粒体P450依赖性单加氧酶系统的功能。