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RIKEN structural genomics beamlines at SPring-8

2002; Wiley; Volume: 58; Issue: s1 Linguagem: Inglês

10.1107/s0108767302097076

1600-5724

Masaki Yamamoto, Takashi Kumasaka, Go Ueno, K. Ida, H. Kanda, Masashi Miyano, T. Ishikawa,

Genomics and Phylogenetic Studies

Protein crystallization is one of the major bottlenecks in protein crystallography.Structural genomics project is expected to be produced a large number of proteins from various organisms every year, and so it will be needed to establish an efficient protocol to obtain suitable crystals for X-ray structural analysis and to develop a full automatic protein crystallization robot.A management system for a large number of data is also indispensable.A challenge to large-scale protein crystallization has started since April 2001 at Highthroughput Factory in RIKEN Harima Institute/SPring-8.In the first step, we made the standardization of protein crystallization procedure: (1) utilizing of a commercially available semi-automatic protein crystallization robot with micro-batch method, (2) establishment of efficient crystallization protocol composed of a set of crystallization screening conditions using each twentyfour precipitants and six buffer solutions, and (3) the evaluation standard of the crystallization-setting scoring.We have performed the preliminary crystallization trials with forty-three proteins from Thermus thermophilus HB8 using these conditions.Crystalline materials obtained from all proteins.Nine of them showed enough diffraction to solve their structures.Based on these results, we have designed and developed full-automatic crystallization and observation robotics, which is provided with liquid-handling and observation procedures for protein crystallization as well as plate stocking.The system can make it easy to manage score record and the date and time of crystallization setup.The plasmids for the protein expression were provided by the RIKEN Structurome Project.