Phase II of the Protein Structure Initiative, funded by the NIH NIGMS (National Institute of General Medical Sciences), is a 5-year effort to determine thousands of protein structures. The New York SGX Research Center for Structural Genomics (NYSGXRC) is one of the four large-scale production centers tasked with determining 100–200 structures annually. Almost all protein production is carried out using the high throughput structural biology platform at SGX Pharmaceuticals (SGX), which supplies 120 or more ultrapure proteins per month for NYSGXRC crystallization and structure determination activities. Protocols for PCR, cloning, expression/solubility testing, fermentation, purification, and crystallization are described. General protocols and detailed experimental results for each target are updated weekly at the public PepcDB website (pepcdb.pdb.org/), and all NYSGXRC clones should be available in 2008 through the PlasmID resource operated by the Harvard Institute of Proteomics.
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Acknowledgments
This work is supported by the Protein Structure Initiative of the National Institute for General Medical Sciences under grant U54-GM74945. The authors thank the members of NYSGXRC for their continued efforts: the SGX informatics, protein production and crystallization teams, the SGX-CAT beamline staff at the APS, the laboratories of Steve Almo at Albert Einstein College of Medicine, S. Swaminathan at Brookhaven National Laboratory, Andrej Sali at University of California San Francisco, and Mark Chance at Case Western Reserve University.
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Sauder, M.J. et al. (2008). High Throughput Protein Production and Crystallization at NYSGXRC. In: Kobe, B., Guss, M., Huber, T. (eds) Structural Proteomics. Methods in Molecular Biology™, vol 426. Humana Press. https://doi.org/10.1007/978-1-60327-058-8_37
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