BIOCHEMICAL AND STRUCTURAL CHARACTERIZATION OF MERCURIC ION REDUCTASE

汞离子还原酶的生化和结构表征

• 批准号: 8363586
• 负责人: Susan Mary Miller
• 金额: $ 1.02万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363586
• 关键词: Alkylmercury lyase; Amino Acids; Binding; Binding Proteins; Biochemical; Catalysis; Catalytic Domain; Chimera organism; Complex; Disulfides; Docking; Drug Metabolic Detoxication; Family; Funding; Glutathione Reductase; Grant; Imagery; Informatics; Ions; Kinetics; Membrane Transport Proteins; Mercury; Metals; Molecular Weight; Mutagenesis; N-terminal; National Center for Research Resources; Oxidoreductase; Pathway interactions; Principal Investigator; Proteins; Research; Research Infrastructure; Resources; Sequence Homology; Site; Source; Structure; Thioredoxin; United States National Institutes of Health; biocomputing; cost; protein protein interaction; pyridine nucleotide

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. This project focuses on the structural and kinetic characterization of mercuric ion reductase. Mercuric ion reductase is composed of two domains, a catalytic core domain that is highly homologous to the FAD containing pyridine nucleotide disulfide reductase family i.e. glutathione reductase and a second domain, a ~69 amino acid N-terminal domain, with sequence homology to metal binding proteins. We are focusing on two aspects: 1) characterization of structural features in the Hg(II) binding pathway that are essential for efficient catalysis and 2) elucidating mechanisms of protein/protein interactions between the catalytic core of MerA and the NmerA domain as well as of core with other proteins of the mercury detoxification pathway including MerB, an organomercurial lyase, and the integral membrane transport proteins, MerT or MerC. We use Chimera to graphically present structures of Hg(II) and other complexes of the catalytic core domain alone and interacting with other small molecular weight proteins like NmerA or thioredoxin. In addition, we use Chimera to assist us in evaluating sites for mutagenesis and examining possible modes for docking between MerA and the other mer pathway proteins.

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。 本项目主要研究汞离子还原酶的结构和动力学特性。汞离子还原酶由两个结构域组成，一个是催化核心区，与含有吡啶核苷酸二硫键还原酶家族的FAD高度同源，即谷胱甘肽还原酶；另一个是第二个结构域，即69个氨基酸的N-末端结构域，序列与金属结合蛋白同源。我们主要集中在两个方面：1)汞(II)结合途径中有效催化所必需的结构特征的表征；2)阐明MERA的催化核心与NmerA结构域以及核心与汞解毒途径的其他蛋白质包括MerB(有机汞裂解酶)和整合膜运输蛋白Mert或Merc之间的蛋白质/蛋白质相互作用的机制。我们使用嵌合体来图形化地显示汞(II)和其他催化核心域的络合物的结构，并与其他小分子蛋白质如NmerA或硫氧还蛋白相互作用。此外，我们使用嵌合体来帮助我们评估突变位点，并检查MERA与其他Mer途径蛋白之间对接的可能模式。