Facilities and methods for the high-throughput crystal structural analysis of human proteins

被引:92
作者
Heinemann, U [1 ]
Büssow, K
Mueller, U
Umbach, P
机构
[1] Max Delbruck Centrum Mol Med, Forsch Grp Kristallog, D-13125 Berlin, Germany
[2] Free Univ Berlin, Inst Chem Kristallog, D-14195 Berlin, Germany
[3] Max Planck Inst Mol Genet, D-14195 Berlin, Germany
[4] Prot Struct Factory, D-14059 Berlin, Germany
[5] BESSY, Prot Struct Factory, D-12489 Berlin, Germany
关键词
D O I
10.1021/ar010129t
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Facilities and methods for the high-throughput crystal structure analysis of human proteins are described as recently established in the Protein Structure Factory, a Berlin-area structural genomics project. Genes encoding human proteins are expressed in either recombinant Escherichia coli or yeast (Saccharomyces cerevisiae or Pichia pastoris). To facilitate and standardize protein purification, the target proteins are produced with various tags for affinity chromatography. For high-throughput crystallization, a robotic station is being set up that has the capacity to handle 960 000 experiments simultaneously. The resulting protein crystals will be subjected to X-ray diffraction experiments at the third-generation synchrotron storage ring BESSY where protein crystallography beamlines are currently under construction. The Protein Structure Factory's strategy for high-throughput production and structure analysis of human proteins is evaluated based on first results.
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收藏
页码:157 / 163
页数:7
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