Facilities and methods for the high-throughput crystal structural analysis of human proteins.
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.
Heinemann
Udo
U
Forschungsgruppe Kristallographie, Max-Delbrück-Centrum für Molekulare Medizin, Robert-Rössle-Strasse 10, D-13125 Berlin, Germany. Heinemann@MDC-Berlin.de
Büssow
Konrad
K
Mueller
Uwe
U
Umbach
Patrick
P
D003001:Cloning, Molecular / Q000379:methods; D018360:Crystallography, X-Ray; D020224:Expressed Sequence Tags; D023281:Genomics / Q000295:instrumentation / Q000379:methods*; D016045:Human Genome Project; D006801:Humans; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry*
D011993:Recombinant Fusion Proteins