STRUCTURAL STUDIES OF MULTICOMPONENT BACTERIAL MONOOXYGENASES

多组分细菌单加氧酶的结构研究

基本信息

  • 批准号:
    7955153
  • 负责人:
  • 金额:
    $ 0.21万
  • 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:
    2009-04-01 至 2010-03-31
  • 项目状态:
    已结题

项目摘要

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bacterial multicomponent monooxygenases are a family of enzymes that utilize a carboxylate-bridged diiron center to hydroxyalate a variety of hydrocarbon substrates. Essential for this activity is the formation of a complex between the hydroxylase and regulatory protein. The structure of the Pseudomonas sp. OX1 phenol hydroxylase, PHH-PHM complex was determined to 2.3 ¿ resolution. The regulatory protein binds on helices A, E, and F of the hydroxylase alpha subunit at the dimer interface 12 ¿ above the diiron center. Although the metal center resembled the structure of mixed-valent methane monooxygenase, significant structural changes were observed in helices E and F that were different than the configuration typically observed in the uncomplexed form of the hydroxylase. These changes have implications for substrate activation and substrate selectivity, the significance of which is currently being explored. Since determining this structure, structural work has focused on obtaining higher resolution data on the PHH-PHM complex with improved regulatory protein occupancy, and crystallographic characterization of various mutants of PHH and toluene/o-xylene monooxygenase hydroxylase from Pseudomonas sp. OX1. Higher resolution and structures of the complex with improved regulatory protein occupancy could confirm mechanistic conclusions and speculations about the structural effects of regulatory protein binding proposed upon analysis of the initial PHH-PHM data. Similarly, a structure of the hydroxylase in the absence of the regulatory protein may reveal critical, mechanistically relevant characteristics of the hydroxylase structure while also providing a platform to further investigate the structural effects of regulatory protein binding on the hydroxylase through comparison with the PHH-PHM structure. Diffraction data from these projects are only preliminary and require improvement and analysis before they may be reported. In effort to obtain a diferric or analogous structure of PHH or PHH-PHM, dithionite soaked divalent metal reconstituted and crystals of both species were pursued but not yet successfully obtained such to yield a quality diffraction data set for structure determination. Mechanistic data on mutant forms of ToMOH (T201X) and PHH (N204X) in which conserved residues near the diiron site were varied by site-directed-mutagenesis indicate roles for these residues, so the mutants were crystallized and their diffraction data collected to yield structural information that complements the mechanistic studies. This structural information is presently being analyzed and prepared for publication in conjuction with the relevant mechanistic studies and their results.
这个子项目是众多研究子项目之一

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

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Stephen J. Lippard其他文献

X-ray structure of a dodecamer duplex containing the major cisplatin d(GpG) intrastrand cross-link
  • DOI:
    10.1016/0162-0134(95)97300-f
  • 发表时间:
    1995-08-01
  • 期刊:
  • 影响因子:
  • 作者:
    Patricia M. Takahara;Amy C. Rosenzweig;Christin A. Frederick;Stephen J. Lippard
  • 通讯作者:
    Stephen J. Lippard
Frank Albert Cotton (1930–2007)
弗兰克·艾伯特·科顿(1930 年至 2007 年)
  • DOI:
    10.1038/446626a
  • 发表时间:
    2007-04-04
  • 期刊:
  • 影响因子:
    48.500
  • 作者:
    Stephen J. Lippard
  • 通讯作者:
    Stephen J. Lippard
High resolution crystal structures of the hydroxylase protein of methane monooxygenase
  • DOI:
    10.1016/0162-0134(95)97479-a
  • 发表时间:
    1995-08-01
  • 期刊:
  • 影响因子:
  • 作者:
    Amy C. Rosenzweig;Pär Nordlund;Stephen J. Lippard;Christin A. Frederick
  • 通讯作者:
    Christin A. Frederick
Conjugués de nanoparticule de polynucléotide polyvalente en tant que véhicules de distribution pour un agent chimiothérapique
多核苷酸多价纳米粒子结合物与化学药物分配载体
  • DOI:
  • 发表时间:
    2010
  • 期刊:
  • 影响因子:
    0
  • 作者:
    Chad A. Mirkin;David A. Giljohann;W. Daniel;Stephen J. Lippard;Shanta Dhar
  • 通讯作者:
    Shanta Dhar
Structural, mechanistic, and model studies for methane monooxygenase
  • DOI:
    10.1016/0162-0134(95)97192-s
  • 发表时间:
    1995-08-01
  • 期刊:
  • 影响因子:
  • 作者:
    Stephen J. Lippard
  • 通讯作者:
    Stephen J. Lippard

Stephen J. Lippard的其他文献

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{{ truncateString('Stephen J. Lippard', 18)}}的其他基金

STRUCTURAL STUDIES OF BACTERIAL MULTICOMPONENT MONOOXYGENASES
细菌多组分单加氧酶的结构研究
  • 批准号:
    8362193
  • 财政年份:
    2011
  • 资助金额:
    $ 0.21万
  • 项目类别:
INVESTIGATIONS OF CISPLATIN-DNA CROSS-LINKS ON NUCLEOSOME CORE PARTICLES
核小体核心颗粒上顺铂-DNA 交联的研究
  • 批准号:
    8169250
  • 财政年份:
    2010
  • 资助金额:
    $ 0.21万
  • 项目类别:
STRUCTURAL STUDIES OF BACTERIAL MULTICOMPONENT MONOOXYGENASES
细菌多组分单加氧酶的结构研究
  • 批准号:
    8170154
  • 财政年份:
    2010
  • 资助金额:
    $ 0.21万
  • 项目类别:
STRUCTURAL STUDIES OF MULTICOMPONENT BACTERIAL MONOOXYGENASES
多组分细菌单加氧酶的结构研究
  • 批准号:
    8169251
  • 财政年份:
    2010
  • 资助金额:
    $ 0.21万
  • 项目类别:
STRUCTURAL STUDIES OF BACTERIAL MULTICOMPONENT MONOOXYGENASES
细菌多组分单加氧酶的结构研究
  • 批准号:
    7954158
  • 财政年份:
    2009
  • 资助金额:
    $ 0.21万
  • 项目类别:
CHEMISTRY AND BIOLOGY OF PLATINUM ANTICANCER DRUGS
铂类抗癌药物的化学和生物学
  • 批准号:
    7955152
  • 财政年份:
    2009
  • 资助金额:
    $ 0.21万
  • 项目类别:
Nonheme Diiron Centers and the Biological Oxidation of Hydrocarbons
非血红素二铁中心和碳氢化合物的生物氧化
  • 批准号:
    7923548
  • 财政年份:
    2009
  • 资助金额:
    $ 0.21万
  • 项目类别:
STRUCTURAL STUDIES OF BACTERIAL MULTICOMPONENT MONOOXYGENASES
细菌多组分单加氧酶的结构研究
  • 批准号:
    7954496
  • 财政年份:
    2009
  • 资助金额:
    $ 0.21万
  • 项目类别:
STRUCTURAL STUDIES OF BACTERIAL MULTICOMPONENT MONOOXYGENASES
细菌多组分单加氧酶的结构研究
  • 批准号:
    7721732
  • 财政年份:
    2008
  • 资助金额:
    $ 0.21万
  • 项目类别:
STRUCTURAL STUDIES OF BACTERIAL MULTICOMPONENT MONOOXYGENASES
细菌多组分单加氧酶的结构研究
  • 批准号:
    7597911
  • 财政年份:
    2007
  • 资助金额:
    $ 0.21万
  • 项目类别:

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