Structure and Function of Integral Membrane Enzyme Human Aromatase

人芳香酶整合膜酶的结构与功能

基本信息

  • 批准号:
    8215735
  • 负责人:
  • 金额:
    $ 40.6万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2009
  • 资助国家:
    美国
  • 起止时间:
    2009-01-01 至 2013-12-31
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): Human cytochrome P450 aromatase (P450arom), an integral membrane hemeprotein of the endoplasmic reticulum, catalyzes the synthesis of estrogens from androgens in the presence of P450-reductase. Despite intense biochemical and biophysical investigations for the past 35 years, the structure-function relationships of P450arom and its catalytic mechanism remain poorly understood. Inhibition of estrogen biosynthesis by P450arom inhibitors is an effective therapy for hormone-dependent breast cancers. Attempts to obtain diffraction-quality crystals of human P450arom have so far been unsuccessful. We have grown single crystals of the androstenedione-complex of the full-length, highly active P450arom purified from human term placenta, gathered complete diffraction data to 2.90E resolution and obtained a solution for the structure. Here, we propose to launch an investigation in order to determine of the structure-function relationships of human P450arom. Our hypothesis is that analysis of the atomic structures of human P450arom-ligand complexes in terms of their functional properties will lead to the elucidation of the origin of substrate and inhibitor specificities, roles of the catalytically important residues, nature of the reaction intermediates, as well as the mechanism of action, and that ligand design and optimization guided by these structural basis will lead to novel high-affinity inhibitors that are exclusive for the target. The specific aims to test the hypothesis are to determine the crystal structures of the complexes of P450arom with its (1) natural substrates androstenedione, testosterone, and 161-hydroxy-testosterone, as well as with (2) the inhibitors exemestane, letrozole, formestane, anastrozole, fadrozole and aminoglutethimide. These findings will reveal the molecular mechanism for substrate and inhibitor specificities and help build a structure-activity database based on the atomic level descriptions of the enzyme-ligand interactions. Next, in specific aim 3 we will investigate the enzyme mechanism by combining the data from aims 1 and 2 with structural data on reaction intermediates. By initiating the aromatization reaction in an enzyme-substrate complex crystal with X-ray photoelectrons during diffraction and in situ monitoring of the Soret band transition with a spectrophotometer, we plan to capture structural snap shots of the reaction intermediates. Completion of these goals will lead to the implementation of specific aim 4 - discovery of new inhibitors through docking, design, synthesis, testing, and optimization, in collaboration with chemists. As a future direction of the project, we plan to crystallize a complex of P450arom with P450- reductase for investigating the mechanism of redox reactions by electron transfer. PUBLIC HEALTH RELEVANCE: Aromatase is a unique enzyme that makes all estrogens in the human body. We propose a research plan to unravel the molecular details of how aromatase works and how aromatase inhibitors prevent it from making estrogens. Results from this investigation will form the basis for future discovery of novel breast cancer drugs that are highly specific for aromatase but cause minimal side effects.
描述(由申请人提供):人细胞色素P450芳香酶(P450 arom)是内质网的一种整合膜血红素蛋白,在P450还原酶存在下催化雄激素合成雌激素。尽管在过去的35年里进行了大量的生物化学和生物物理研究,但P450 arom的结构-功能关系及其催化机制仍然知之甚少。通过P450 arom抑制剂抑制雌激素生物合成是治疗乳腺癌的有效方法。迄今为止,获得衍射质量的人类P450 arom晶体的尝试都没有成功。我们已经生长了从人足月胎盘中纯化的全长、高活性P450 arom的雄烯二酮复合物的单晶,收集了完整的衍射数据至2.90E分辨率,并获得了结构的解决方案。在这里,我们建议开展调查,以确定人类P450 arom的结构与功能的关系。我们的假设是,根据其功能特性分析人P450芳族配体复合物的原子结构将导致阐明底物和抑制剂特异性的起源、催化重要残基的作用、反应中间体的性质以及作用机制,并且由这些结构基础指导的配体设计和优化将导致对靶专一的新型高亲和力抑制剂。检验该假设的具体目的是确定P450 arom与其(1)天然底物雄烯二酮、睾酮和161-羟基-睾酮以及(2)抑制剂阿司美坦、来曲唑、福美坦、阿那曲唑、法倔唑和氨鲁米特的复合物的晶体结构。这些发现将揭示底物和抑制剂特异性的分子机制,并有助于建立基于原子水平描述的酶-配体相互作用的结构-活性数据库。接下来,在具体目标3中,我们将通过结合目标1和2的数据与反应中间体的结构数据来研究酶的机制。通过在衍射过程中用X射线光电子在酶-底物复合物晶体中引发芳构化反应,并用分光光度计原位监测Soret带跃迁,我们计划捕获反应中间体的结构快照。这些目标的完成将导致具体目标4的实施-通过对接,设计,合成,测试和优化,与化学家合作,发现新的抑制剂。作为该项目的未来发展方向,我们计划结晶P450芳香族化合物与P450还原酶的复合物,以研究电子转移氧化还原反应的机理。公共卫生相关性:芳香酶是一种独特的酶,使所有雌激素在人体内。我们提出了一个研究计划,以解开芳香化酶如何工作的分子细节,以及芳香化酶抑制剂如何阻止它产生雌激素。这项研究的结果将成为未来发现新型乳腺癌药物的基础,这些药物对芳香酶具有高度特异性,但副作用最小。

项目成果

期刊论文数量(10)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Novel aromatase inhibitors by structure-guided design.
  • DOI:
    10.1021/jm300930n
  • 发表时间:
    2012-10-11
  • 期刊:
  • 影响因子:
    7.3
  • 作者:
    Ghosh, Debashis;Lo, Jessica;Morton, Daniel;Valette, Damien;Xi, Jingle;Griswold, Jennifer;Hubbell, Susan;Egbuta, Chinaza;Jiang, Wenhua;An, Jing;Davies, Huw M. L.
  • 通讯作者:
    Davies, Huw M. L.
Motion and flexibility in human cytochrome p450 aromatase.
  • DOI:
    10.1371/journal.pone.0032565
  • 发表时间:
    2012
  • 期刊:
  • 影响因子:
    3.7
  • 作者:
    Jiang W;Ghosh D
  • 通讯作者:
    Ghosh D
Convenient method for the functionalization of the 4- and 6-positions of the androgen skeleton.
Recent Progress in the Discovery of Next Generation Inhibitors of Aromatase from the Structure-Function Perspective.
  • DOI:
    10.1021/acs.jmedchem.5b01281
  • 发表时间:
    2016-06-09
  • 期刊:
  • 影响因子:
    7.3
  • 作者:
    Ghosh D;Lo J;Egbuta C
  • 通讯作者:
    Egbuta C
{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

DEBASHIS GHOSH其他文献

DEBASHIS GHOSH的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('DEBASHIS GHOSH', 18)}}的其他基金

INTEGRAL MEMBRANE ENZYMES IN ESTROGEN BIOSYNTHESIS
雌激素生物合成中的整体膜酶
  • 批准号:
    8363523
  • 财政年份:
    2011
  • 资助金额:
    $ 40.6万
  • 项目类别:
INTEGRAL MEMBRANE ENZYMES IN ESTROGEN BIOSYNTHESIS
雌激素生物合成中的整体膜酶
  • 批准号:
    8171503
  • 财政年份:
    2010
  • 资助金额:
    $ 40.6万
  • 项目类别:
Structure and Function of Integral Membrane Enzyme Human Aromatase
人芳香酶整合膜酶的结构与功能
  • 批准号:
    7743555
  • 财政年份:
    2009
  • 资助金额:
    $ 40.6万
  • 项目类别:
Structure and Function of Integral Membrane Enzyme Human Aromatase
人芳香酶整合膜酶的结构与功能
  • 批准号:
    8208078
  • 财政年份:
    2009
  • 资助金额:
    $ 40.6万
  • 项目类别:
INTEGRAL MEMBRANE ENZYMES IN ESTROGEN BIOSYNTHESIS
雌激素生物合成中的整体膜酶
  • 批准号:
    7955568
  • 财政年份:
    2009
  • 资助金额:
    $ 40.6万
  • 项目类别:
Structure and Function of Integral Membrane Enzyme Human Aromatase
人芳香酶整合膜酶的结构与功能
  • 批准号:
    8189257
  • 财政年份:
    2009
  • 资助金额:
    $ 40.6万
  • 项目类别:
CRYSTALS OF 143 KDA BOVINE INTERPHOTORECEPTOR RETINOID BINDING PRO
143 KDA 牛中间光感受器视黄醇结合 PRO 的晶体
  • 批准号:
    7721299
  • 财政年份:
    2008
  • 资助金额:
    $ 40.6万
  • 项目类别:
INTEGRAL MEMBRANE ENZYMES IN ESTROGEN BIOSYNTHESIS
雌激素生物合成中的整体膜酶
  • 批准号:
    7721336
  • 财政年份:
    2008
  • 资助金额:
    $ 40.6万
  • 项目类别:
CRYSTALS OF 143 KDA BOVINE INTERPHOTORECEPTOR RETINOID BINDING PRO
143 KDA 牛中间光感受器视黄醇结合 PRO 的晶体
  • 批准号:
    7598553
  • 财政年份:
    2007
  • 资助金额:
    $ 40.6万
  • 项目类别:
INTEGRAL MEMBRANE ENZYMES IN ESTROGEN BIOSYNTHESIS
雌激素生物合成中的整体膜酶
  • 批准号:
    7598554
  • 财政年份:
    2007
  • 资助金额:
    $ 40.6万
  • 项目类别:

相似海外基金

Collaborative Research: Beyond the Single-Atom Paradigm: A Priori Design of Dual-Atom Alloy Active Sites for Efficient and Selective Chemical Conversions
合作研究:超越单原子范式:双原子合金活性位点的先验设计,用于高效和选择性化学转化
  • 批准号:
    2334970
  • 财政年份:
    2024
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Standard Grant
NSF-BSF: Towards a Molecular Understanding of Dynamic Active Sites in Advanced Alkaline Water Oxidation Catalysts
NSF-BSF:高级碱性水氧化催化剂动态活性位点的分子理解
  • 批准号:
    2400195
  • 财政年份:
    2024
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Standard Grant
Collaborative Research: Beyond the Single-Atom Paradigm: A Priori Design of Dual-Atom Alloy Active Sites for Efficient and Selective Chemical Conversions
合作研究:超越单原子范式:双原子合金活性位点的先验设计,用于高效和选择性化学转化
  • 批准号:
    2334969
  • 财政年份:
    2024
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Standard Grant
Mechanochemical synthesis of nanocarbon and design of active sites for oxygen reducton/evolution reactions
纳米碳的机械化学合成和氧还原/演化反应活性位点的设计
  • 批准号:
    23K04919
  • 财政年份:
    2023
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Creation of porous inorganic frameworks with controlled structure of metal active sites by the building block method.
通过积木法创建具有金属活性位点受控结构的多孔无机框架。
  • 批准号:
    22KJ2957
  • 财政年份:
    2023
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Grant-in-Aid for JSPS Fellows
Catalysis of Juxaposed Active Sites Created in Nanospaces and Their Applications
纳米空间中并置活性位点的催化及其应用
  • 批准号:
    23K04494
  • 财政年份:
    2023
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Generation of carbon active sites by modifying the oxygen containing functional groups and structures of carbons for utilizing to various catalytic reactions.
通过修饰碳的含氧官能团和结构来产生碳活性位点,用于各种催化反应。
  • 批准号:
    23K13831
  • 财政年份:
    2023
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Grant-in-Aid for Early-Career Scientists
CAREER: CAS: Understanding the Chemistry of Palladium and Silyl Compounds to Design Catalyst Active Sites
职业:CAS:了解钯和甲硅烷基化合物的化学性质以设计催化剂活性位点
  • 批准号:
    2238379
  • 财政年份:
    2023
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Continuing Grant
CAS: Collaborative Research: Tailoring the Distribution of Transient vs. Dynamic Active Sites in Solid-Acid Catalysts and Their Impacts on Chemical Conversions
CAS:合作研究:定制固体酸催化剂中瞬时活性位点与动态活性位点的分布及其对化学转化的影响
  • 批准号:
    2154399
  • 财政年份:
    2022
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Standard Grant
Engineering of Active Sites in Heterogeneous Catalysts for Sustainable Chemical and Fuel Production.
用于可持续化学和燃料生产的多相催化剂活性位点工程。
  • 批准号:
    RGPIN-2019-06633
  • 财政年份:
    2022
  • 资助金额:
    $ 40.6万
  • 项目类别:
    Discovery Grants Program - Individual
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了