METHANOCALDOCOCCUS JANNASCHII COBY (MJ1117)

甲烷热球菌 JANNASCHII COBY (MJ1117)

• 批准号: 7954616
• 负责人: JORGE C ESCALANTE
• 金额: $ 0.16万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2010-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954616
• 关键词: Anabolism; Archaea; Binding; Biochemical; Carbon; Characteristics; Cobalamin; Computer Retrieval of Information on Scientific Projects Database; Enzymes; Escherichia coli; Funding; Genes; Grant; Guanosine Triphosphate; Institution; Kinetics; Label; NMR Spectroscopy; Nitrogen; Nuclear Magnetic Resonance; Phosphotransferases; Proteins; Reporting; Research; Research Personnel; Resources; Salmonella enterica; Solutions; Source; Specificity; Structure; United States National Institutes of Health; base; guanylyltransferase; inorganic phosphate; overexpression; three dimensional structure

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. Structural and Biochemical Characterization of the GTP:Adenosylcobinamide-phosphate Guanylyltransferase (CobY) Enzyme from the Hyperthermophilic Archaeon Methanocaldococcus jannaschii The archaeal cobY gene is the nonorthologous replacement of the bacterial NTP:adenosylcobinamide kinase/GTP:adenosylcobinamide-phosphate guanylyltransferase (cobU in Salmonella enterica) and is required for the biosynthesis of cobalamin. We overexpressed the Methanocaldococcus jannaschii MJ1117 (cobY) gene product (Mj CobY) in Escherichia coli and purified it to homogeneity. We report here the three-dimensional structure of Mj CobY, determined by solution nuclear magnetic resonance (NMR) spectroscopy from protein labeled with carbon-13 and nitrogen-15. We show that Mj CobY has the GTP:adenosylcobinamide-phosphate guanylyltransferase activity associated with CobU. The biochemical characterization of Mj CobY includes an analysis of substrate binding and specificity, physical characteristics, and kinetic parameters. Based on the Mj CobY structure and biochemical parameters, we propose a mechanism whereby Mj CobY transferse the guanylyl moiety of GTP to adenosylcobinamide-phosphate during the synthesis of cobalamin.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 Structural and Biochemical Characterization of the GTP:Adenosylcobinamide-phosphate Guanylyltransferase (CobY) Enzyme from the Hyperthermophilic Archaeon Methanocaldococcus jannaschii The archaeal cobY gene is the nonorthologous replacement of the bacterial NTP:adenosylcobinamide激酶/GTP：腺基辛酰胺 - 磷酸圭尼甘油酯转移酶（肠道沙门氏菌中的COBU），是钴胺素的生物合成所必需的。 我们过度表达了jannaschii MJ1117（Coby）基因产物（MJ Coby）的大肠杆菌，并将其纯化为同质性。 我们在这里报告了MJ Coby的三维结构，该结构由用碳-13和氮15标记的蛋白质的溶液核磁共振（NMR）光谱确定。 我们表明，MJ Coby具有GTP：与COBU相关的腺苷酰胺 - 磷酸胰鸟酰胺胰岛转移酶活性。 MJ Coby的生化特征包括对底物结合和特异性，物理特征和动力学参数的分析。 基于MJ Coby结构和生化参数，我们提出了一种机制，通过该机制，MJ Coby在合成钴胺素期间将GTP转移到Guanylyl部分到腺苷酸磷酸盐。