CRYSTAL STRUCTURE OF E COLI TRANSHYDROGENASE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 在这个新的、单一的实验方案中，需要光束时间来筛选大肠杆菌转氢酶(TH)的晶体进行衍射，并收集自然数据。TH是一种206 kDa的完整膜蛋白，是维持线粒体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。实验将需要在筛选低温保护条件的同时，评估由多种TH结构和各种洗涤剂存在的情况下制备的晶体形态。一旦建立了适度的衍射，将收集用于分子置换计算的自然数据，使用已知的TH可溶域的晶体结构。如果这些都不成功，将采用确定相的经验方法。

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.