CRYSTAL STRUCTURE OF E COLI TRANSHYDROGENASE

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 在这个新的单一实验方案中，需要束流时间来筛选E.大肠杆菌转氢酶（TH）的衍射和收集天然数据。TH是一种206 kDa的整合膜蛋白，是维持线粒体中NADPH水平所必需的构象偶联蛋白泵。TH突变与2型糖尿病有关。实验将需要评估从多种TH构建体制备的晶体形式，并在各种洗涤剂的存在下，同时筛选冷冻保护条件。一旦建立了中等衍射，将使用TH可溶结构域的已知晶体结构收集天然数据用于分子置换计算。如果这些方法不成功，则将采用经验性的相位确定方法。

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.