Development of X-ray intensity data-collection system for macromolecular crystals

研制了一套实验室生物大分子晶体X射线强度数据采集系统。该系统包括一个新的X射线探测器（RAXIS-IV）和一个改进的细聚焦（0.1 mm x 1.0 mm）X射线发生器（FR-C）。使用两个大的（300 mm x 300 mm）成像板，检测器的设计产生一个高灵敏度的短读出时间。改进了X射线发生器的油密封和真空密封，以产生持久的密封。X射线束用Supper的双镜聚焦。该系统被用于几个蛋白质-DNA复合物，蛋白质-GTP复合物和蛋白质晶体的数据收集和这些晶体的结构测定进行使用所获得的强度数据。它们是与DNA复合的芽殖酵母转录因子PHO 4、与DNA复合的小鼠转录因子IRF-2、与DNA复合的裂殖酵母转录因子Pap 1、小鼠海马丝氨酸蛋白酶neuropsin、与GTP γ S复合的人小G蛋白RhoA、与GTP γ S复合的人小G蛋白RhoA、与大肠杆菌应答调节剂CheY复合的大肠杆菌传感激酶ArcB的HPt结构域、ArcB的HPt结构域是大肠杆菌磷酸组氨酸磷酸酶SixA。

An instrument system for X-ray intensity data collection from bio-macromolecular crystals in a laboratory was developed. The system consists of a new X-ray detector (RAXIS-IV) and an improved fine focused (0.1 mm x 1.0 mm) X-ray generator (FR-C). Using two large (300 mm x 300 mm) imaging plates, the detector was designed to yield a short read-out time with high sensitivity. The oil and vacuum sealing of the X-ray generator was improved to yield long-lasting seals. The X-ray beam was focused with Supper's double-mirror. The system was used for data collections of several protein-DNA complex, protein-GTP complex and protein crystals and structure determinations of these crystals were performed using the intensity data obtained. These are the budding yeast transcription factor PHO4 complexed with DNA,the mouse transcription factor IRF-2 complexed with DNA,the fission yeast transcription factor Pap1 complexed with DNA,mouse hippocampal serine protease neuropsin, the human small G protein RhoA complexed with GTPgammaS,the human small G protein RhoA complexed with GTPgammaS,the HPt domain of a E.coli sensor kinase ArcB complexed with a E.coli response regulator CheY,the HPt domain of ArcB a E.coli phospho-histidine phosphatase SixA.