Sugahara Michihiro, Asada Yukuhiko, Shimizu Katsumi, Yamamoto Hitoshi, Lokanath Neratur K, Mizutani Hisashi, Bagautdinov Bagautdin, Matsuura Yoshinori, Taketa Midori, Kageyama Yuichi, Ono Naoko, Morikawa Yuko, Tanaka Yukiko, Shimada Hiroki, Nakamoto Takanobu, Sugahara Mitsuaki, Yamamoto Masaki, Kunishima Naoki
Advanced Protein Crystallography Research Group, RIKEN SPring-8 Center, Harima Institute, 1-1-1 Kouto, Sayo-cho, Sayo-gun, Hyogo, 679-5148, Japan.
J Struct Funct Genomics. 2008 Dec;9(1-4):21-8. doi: 10.1007/s10969-008-9042-y. Epub 2008 Aug 2.
A high-throughput crystallization-to-structure pipeline for structural genomics was recently developed at the Advanced Protein Crystallography Research Group of the RIKEN SPring-8 Center in Japan. The structure determination pipeline includes three newly developed technologies for automating X-ray protein crystallography: the automated crystallization and observation robot system "TERA", the SPring-8 Precise Automatic Cryosample Exchanger "SPACE" for automated data collection, and the Package of Expert Researcher's Operation Network "PERON" for automated crystallographic computation from phasing to model checking. During the 5 years following April, 2002, this pipeline was used by seven researchers to determine 138 independent crystal structures (resulting from 437 purified proteins, 234 cryoloop-mountable crystals, and 175 diffraction data sets). The protocols used in the high-throughput pipeline are described in this paper.
日本理化学研究所SPring-8中心的高级蛋白质晶体学研究小组最近开发了一种用于结构基因组学的高通量从结晶到结构解析流程。该结构解析流程包括三项新开发的用于自动化X射线蛋白质晶体学的技术:自动化结晶和观察机器人系统“TERA”、用于自动数据收集的SPring-8精确自动低温样品交换器“SPACE”以及用于从相位分析到模型检查的自动化晶体学计算的专家研究员操作网络包“PERON”。在2002年4月之后的5年里,七名研究人员使用该流程确定了138个独立的晶体结构(来自437种纯化蛋白质、234个可通过低温环安装的晶体和175个衍射数据集)。本文描述了高通量流程中使用的实验方案。