CRYSTAL STRUCTURE OF E COLI TRANSHYDROGENASE

大肠杆菌转氢酶的晶体结构

• 批准号: 8170092
• 负责人: Charles David Stout
• 金额: $ 0.37万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170092
• 关键词: Computer Retrieval of Information on Scientific Projects Database; Coupled; Data; Detergents; Escherichia coli; Funding; Grant; Institution; Integral Membrane Protein; Link; Maintenance; Methods; Mitochondria; Molecular; Mutation; NAD(P)+ transhydrogenase; NADP; Non-Insulin-Dependent Diabetes Mellitus; Phase; Proteins; Pump; Research; Research Personnel; Resources; Screening procedure; Source; Structure; Time; United States National Institutes of Health; research study

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. In this new, single experiment proposal, beam time is requested to screen crystals of E. coli transhydrogenase (TH) for diffraction and to collect native data. TH is a 206 kDa integral membrane protein and a conformationally coupled protein pump essential for the maintenance of NADPH levels in mitochondria. TH mutations are linked to Type 2 diabetes. Experiments will entail evaluating crystal forms prepared from multiple TH constructs and in the presence of various detergents while screening cryoprotection conditions. Once moderate diffraction is established, native data will be collected for molecular replacement calculations, using known crystal structures of TH soluble domains. If these are unsuccessful, empirical methods of phase determination will be employed.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 在这个新的单一实验提案中，需要光束时间来筛选大肠杆菌转氢酶 (TH) 晶体的衍射并收集原始数据。 TH 是一种 206 kDa 的整合膜蛋白和构象耦合蛋白泵，对于维持线粒体中的 NADPH 水平至关重要。 TH 突变与 2 型糖尿病有关。实验将需要评估由多种 TH 构建体制备的晶体形式，并在各种去污剂存在下，同时筛选冷冻保护条件。一旦建立了适度的衍射，将使用 TH 可溶域的已知晶体结构收集原始数据用于分子替换计算。如果这些不成功，将采用相位确定的经验方法。