CYP17A1 STRUCTURE FUNCTION, CRITICAL ENZYME IN HUMAN ANDROGEN BIOSYNTHESIS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 前列腺癌是美国最常见的癌症，也是男性癌症相关死亡的第二大原因。进展与雄激素的存在密切相关。传统的雄激素剥夺疗法显著减少了前列腺组织中雄激素的产生，但未能抑制肾上腺雄激素的合成，从而使血液中的雄激素水平保持在基础水平。细胞色素P450 17A1(CYP17A1)负责在这两种组织中将孕烯醇酮转化为雄激素。因此，在前列腺癌的治疗中，抑制CYP17A1为完全去除雄激素提供了一种策略。几种CYP17A1抑制剂目前正在进行临床试验，但目前还没有关于CYP17A1的结构信息。由于缺乏CYP17A1晶体结构，因此无法理解目前的抑制剂是如何起作用的，以及如何改进它们。这项建议的目的是确定CYP17A1的结构。了解CYP17A1与其配体之间的相互作用将提供有关该酶如何发挥作用的信息。我们的中心假设是，CYP17A1的晶体结构将揭示与酶的独特特征相对应的特定相互作用。其具体目标是：1)设计一种可溶版本的CYP17A1，2)优化表达和纯化策略，以产生大量纯的、稳定的、功能活性的、单分散的适合结晶的CYP17A1，以及3)生长衍射级的CYP17A1晶体。拟议研究的预期结果是获得重要的类固醇生物合成酶CYP17A1的第一个晶体结构。这种结构将使我们能够详细地了解配体结合模式。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Prostate cancer is the most common occurring cancer and the second-leading cause of cancer-related death for men in the United States. Progression is strongly linked to the presence of androgens. Traditional androgen deprivation therapy significantly decreases androgen production in prostate tissue, but fails to inhibit adrenal androgen synthesis, leaving a basal level of androgens in circulation. Cytochrome P450 17A1 (CYP17A1) is responsible for the conversion of pregnenolone to androgens in both tissues. Thus, inhibition of CYP17A1 provides a strategy for complete deprivation of androgens in the treatment of prostate cancer. Several CYP17A1 inhibitors are currently in clinical trials, but there is currently no structural information about CYP17A1. The lack of a CYP17A1 crystal structure prevents an understanding of how current inhibitors work and how they might be improved. The objective of this proposal is to determine a structure of CYP17A1. Understanding the interaction between CYP17A1 and its ligands would provide information about how this enzyme functions. Our central hypothesis is that a crystal structure of CYP17A1 will reveal specific interactions that correspond to unique features of the enzyme. The specific aims are: 1) to engineer a soluble version of CYP17A1, 2) to optimize expression and purification strategies to generate mg quantities of pure, stable, functionally active, monodisperse CYP17A1 suitable for crystallization efforts, and 3) to grow diffraction-quality crystals of CYP17A1. The expected outcome of the proposed study is the first crystal structure of the important steroid biosynthetic enzyme CYP17A1. This structure will enable a detailed understanding of ligand binding modes.