Dynamics and Interactions of Cytochrome P450 19A1

细胞色素 P450 19A1 的动力学和相互作用

• 批准号: 10201672
• 负责人: John C Hackett
• 金额: $ 7.6万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2021-08-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10201672
• 关键词: Address; Adopted; Adverse effects; Architecture; Aromatase; Binding Proteins; Binding Sites; Catalysis; Clinic; Complement; Complex; Cytochrome P450; Cytochrome a; Data; Deuterium; Disease; Drug Interactions; Electron Transport; Endocrine System Diseases; Environment; Enzymes; Equilibrium; Estrogens; Estrone; Family; Heme; Hormones; Human; Hybrids; Hydrogen; Infertility; Investigation; Kinetics; Ligand Binding; Ligands; Lipid Bilayers; Lipids; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Membrane; Methods; Modeling; Neutrons; Oxidation-Reduction; Pathway interactions; Pharmaceutical Preparations; Pharmacotherapy; Phylogenetic Analysis; Physiological; Population; Positioning Attribute; Property; Protein Conformation; Protein Dynamics; Proteins; Public Health; Research; Resources; Roentgen Rays; Role; Sampling; Structure; Surface; Testing; Tissues; Trees; Variant; Xenobiotics; anastrozole; androgen biosynthesis; conformational conversion; drug development; flexibility; inhibitor/antagonist; insight; malignant endocrine gland neoplasm; molecular dynamics; nanodisk; novel; novel strategies; preference; steroid hormone; tool

The cytochromes P450 (CYPs) are responsible for a dazzling array of transformations in the disposition of xenobiotics and biosyntheses of hormones. Over 70% of drugs are metabolized by CYPs; hence tremendous resources are devoted to identification of substrates and inhibitors in drug development to avoid later attrition, or adverse drug interactions following introduction into the clinic. Among these, CYPs involved in steroid hormone biosyntheses are among the key targets for the pharmacotherapy of endocrine disorders and cancers. The range of catalytic selectivity and the breadth of substrates transformed by CYPs are immense; however the interplay of protein dynamics, ligand binding, and catalysis used to achieve this flexibility remains poorly understood. Within this void, nothing is known about how these are impacted by their physiological context. We have adopted human aromatase (CYP19A1), responsible for estrogen biosynthesis from androgens, as a model to address these longstanding and important questions. Phylogenetic analysis supports that CYP19A1 is among the most primordial human CYPs and it is functionally representative of those steroidogenic enzymes catalyzing sequential transformations. The translational relevance of CYP19A1 cannot be underestimated since it has vital role in maintaining numerous tissues and has demonstrated immense value as a pharmacotherapy target for gynecological disorders, cancers and infertility. There are three Specific Aims to test our hypotheses, we will: 1) Map pathways for 19A1 ligand entry, product egress, and delineate the structural underpinnings of selectivity. The working hypotheses are that i) substrates, intermediate products, and inhibitors give rise to unique equilibrium dynamics and ii) that selectivity is conferred by transient bound states as ligands enter and exit the enzyme through distinct channels. 2) Delineate the impact of membrane complexity on catalysis, ligand binding kinetics and global dynamics in 19A1. The working hypotheses are i) Selectivity is dominated by kinetics involving multiple protein states and conformational transitions, ii) catalytic and inhibitory 19A1 complexes display distinct dynamics, and iii) both protein dynamics and binding kinetics are impacted by membrane properties. 3) Determine the architecture of 19A1-NDs in variable environments by small-angle x-ray and neutron scattering. The working hypothesis is that the insertion depth and orientation of 19A1 are determined by the surface and bulk properties of the bilayer. These Aims are technically- independent, but convergent approaches that are highly synergistic in combination, and will not only allow novel insight into CYP19A1, but also unveil paradigms that will transform our understanding of CYP function.

细胞色素P450(Cyps)负责一系列令人眼花缭乱的转化 外源物质和激素的生物合成。超过70%的药物是由Cyps代谢的；因此 资源用于药物开发中的底物和抑制剂的识别，以避免以后的磨损， 或药物进入临床后的不良反应。其中，Cyps与类固醇有关 激素生物合成是内分泌疾病药物治疗的关键靶点之一， 癌症。其催化选择性范围和转化底物的广度很大； 然而，用于实现这种灵活性的蛋白质动力学、配体结合和催化的相互作用仍然存在 人们对此知之甚少。在这一空白中，我们对这些如何受到生理上的影响一无所知。 背景。我们采用了人类芳香酶(CYP19A1)，负责从 雄激素，作为解决这些长期而重要的问题的模型。系统发育分析支持 CyP19A1是人类最原始的Cyps之一，它在功能上代表了这些 类固醇生成酶催化顺序转化。CYP19A1的翻译关联性不能 被低估，因为它在维持众多组织方面具有至关重要的作用，并已显示出巨大的 作为妇科疾病、癌症和不孕症的药物治疗靶点的价值。有三个具体的 为了验证我们的假设，我们将：1)绘制19A1配体进入、产物出口的路径图，并描绘出 选择性的结构性基础。工作假设是i)底物，中间产物， 而抑制剂产生了独特的平衡动力学，以及ii)选择性由瞬时结合所赋予 以配体的形式通过不同的通道进入和离开酶。2)描绘薄膜的影响 19A1关于催化、配体结合动力学和全球动力学的复杂性。工作假设为i) 选择性由涉及多个蛋白质状态和构象转变的动力学控制，II)催化 和抑制性19A1复合体表现出明显的动力学特征，以及III)蛋白质动力学和结合动力学 会受到膜性能的影响。3)通过以下方式确定19A1-NDS在可变环境中的体系结构 小角x射线和中子散射。工作假设是插入深度和方向 19A1由双层的表面和体相性质决定。这些目标从技术上讲是- 独立但融合的方法，在组合中高度协同，不仅将允许 对CYP19A1的新见解，但也揭示了将改变我们对CYP功能的理解的范式。