Structure and Function of P450 Enzymes in Steroid Hormone Biosynthesis

类固醇激素生物合成中 P450 酶的结构和功能

基本信息

批准号：
9130194
负责人：
MARTIN EGLI
金额：
$ 41.13万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-30 至 2018-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9130194
关键词：
Active Sites Address Adrenal Cortex Amino Acids Anabolism Aryl Hydrocarbon Hydroxylases Beryllium Binding Biochemical Biological Carbon Carcinogens Cattle Clinical Congenital adrenal hyperplasia Cytochrome P450 Cytochromes Cytochromes b5 Disease Eicosanoids Elements Endocrine System Diseases Enzymes Genes Genetic Glucocorticoids Goals Gonadal Steroid Hormones Gonadal structure Health Heme Hormones Human Human Activities Hydroxylation Indium Individual Kinetics Lead Location Malignant neoplasm of prostate Methodology Mineralocorticoids Mixed Function Oxygenases Mutagenesis Mutation Oxidoreductase Pathway interactions Pharmaceutical Preparations Play Positioning Attribute Pregnenolone Progesterone Property Proteins Reaction Reporting Resolution Roentgen Rays Role Series Site Site-Directed Mutagenesis Steroid 17-alpha-monooxygenase Steroid 21-Monooxygenase Steroid Hydroxylases Steroids Structure Structure-Activity Relationship Time Tissues Variant X-Ray Crystallography Xenobiotic Metabolism Zebrafish base cell type drug candidate hormone biosynthesis inhibitor/antagonist innovation insight interest member mutant novel protein expression protein function protein structure research study steroid hormone steroid hormone biosynthesis targeted treatment

项目摘要

DESCRIPTION (provided by applicant): The large cytochrome P450 (P450, CYP) superfamily is distributed throughout all biological kingdoms. The two general classes of P450 reactions are xenobiotic metabolism (e.g., drugs, carcinogens) and endogenous substrate biosynthesis (e.g., hormones, eicosanoids). High-resolution structures of members of ~50 subfamilies among the >17,000 P450 superfamily members are known (all showing a common structural fold), but the precise roles of most individual structural elements are not clearly established. We propose to address this topic by investigating two P450s involved in steroid hormone biosynthesis: mammalian P450 21A2, which is known to have >100 amino acid mutations that influence its steroid 21-hydroxylation activity, and human P450 17A1 and zebrafish P450s 17A1 and 17A2, the latter two being distinct enzymes with dramatic variations in their steroid 17α-hydroxylation/17,20-lyase activities. By analyzing these enzymes in detail (structure and kinetics), we will be able to establish a much more precise view of P450 structure/function than is currently available. We anticipate that these studies will clarify in detail how these two important P450s can function in steroid hormone biosynthesis, and the results can then be applied to understanding the structure/function of other P450s. Mutations in both P450 21A2 and 17A1 are the major causes of a genetic group of diseases known as congenital adrenal hyperplasia, and P450 17A1 is an important drug target for treatment of prostate cancer. The results from these studies will allow a better understanding of both diseases. Taking advantage of determining how amino acid alterations modify functional properties of these enzymes, we will identify the roles that specific structural regions play. The methodology will involve heterologous protein expression/purification/mutagenesis, X-ray crystallography, detailed kinetic analysis, and other biochemical studies (e.g., binding of substrates and other proteins). It is expected that these innovative studies will produce novel insights into and unexpected new understanding of P450 structure/function.

描述（由适用提供）：大细胞色素P450（P450，CYP）超家族均分布在所有生物学界。 P450的两种一般类别是异生元代谢（例如，药物，癌剂）和内源性底物生物合成（例如激素，eicosanoids）。 > 17,000个P450超家族成员之间约50个亚家族成员的高分辨率结构是已知的（均显示出常见的结构折叠），但是大多数单个结构元素的确切作用尚未明确确定。我们建议通过调查参与类固醇马酮生物合成的两种P450：哺乳动物P450 21A2，已知它具有> 100个氨基酸突变，这些氨基酸突变影响其类固醇21-羟基化活性，人类P450 17a1和Zebrafish P450S 17A1和17a2，seltiatiation seltiatiation seltiatiation seltiatiation in sertiatiation ins seltiatiation 17α-羟基化/17,20-裂解活性。通过详细分析这些酶（结构和动力学），我们将能够建立比当前可用的P450结构/功能更精确的视图。我们预计这些研究将详细阐明这两种重要的P450如何在类类赛马的生物合成中起作用，然后可以将结果应用于理解其他P450的结构/功能。 P450 21a2和17a1中的突变是一种称为先天性肾上腺增生的遗传疾病的主要原因，P450 17A1是治疗前列腺癌的重要药物靶标。这些研究的结果将可以更好地了解这两种疾病。利用确定氨基酸的改变如何改变这些酶的功能特性，我们将确定特定结构区域的作用。该方法将涉及异源蛋白表达/纯化/诱变，X射线晶体学，详细的动力学分析以及其他生化研究（例如底物和其他蛋白质的结合）。预计这些创新的研究将产生对P450结构/功能的新见解和意想不到的新理解。

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Functional analysis of human cytochrome P450 21A2 variants involved in congenital adrenal hyperplasia.

参与先天性肾上腺增生的人细胞色素 P450 21A2 变体的功能分析。

DOI：
10.1074/jbc.m117.792465
发表时间：
2017
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Wang,Chunxue;Pallan,PradeepS;Zhang,Wei;Lei,Li;Yoshimoto,FrancisK;Waterman,MichaelR;Egli,Martin;Guengerich,FPeter
通讯作者：
Guengerich,FPeter

In Situ Proteolysis for Crystallization of Membrane Bound Cytochrome P450 17A1 and 17A2 Proteins from Zebrafish.