Structural genomics of the Thermotoga maritima proteome implemented in a high-throughput structure determination pipeline

Scott A. Lesley, Peter Kuhn, Adam Godzik, Ashley M. Deacon, Irimpan Mathews, Andreas Kreusch, Glen Spraggon, Heath E. Klock, Daniel McMullan, Tanya Shin, Juli Vincent, Alyssa Robb, Linda S. Brinen, Mitchell D. Miller, Timothy M. McPhillips, Mark A. Miller, Daniel Scheibe, Jaume M. Canaves, Chittibabu Guda, Lukasz JaroszewskiThomas L. Selby, Marc Andre Elsliger, John Wooley, Susan S. Taylor, Keith O. Hodgson, Ian A. Wilson, Peter G. Schultz, Raymond C. Stevens

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373 Scopus citations

Abstract

Structural genomics is emerging as a principal approach to define protein structure-function relationships. To apply this approach on a genomic scale, novel methods and technologies must be developed to determine large numbers of structures. We describe the design and implementation of a high-throughput structural genomics pipeline and its application to the proteome of the thermophilic bacterium Thermotoga maritima. By using this pipeline, we successfully cloned and attempted expression of 1,376 of the predicted 1,877 genes (73%) and have identified crystallization conditions for 432 proteins, comprising 23% of the T. maritima proteome. Representative structures from TM0423 glycerol dehydrogenase and TM0449 thymidylate synthase-complementing protein are presented as examples of final outputs from the pipeline.

Original languageEnglish (US)
Pages (from-to)11664-11669
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume99
Issue number18
DOIs
StatePublished - Sep 3 2002

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    Lesley, S. A., Kuhn, P., Godzik, A., Deacon, A. M., Mathews, I., Kreusch, A., Spraggon, G., Klock, H. E., McMullan, D., Shin, T., Vincent, J., Robb, A., Brinen, L. S., Miller, M. D., McPhillips, T. M., Miller, M. A., Scheibe, D., Canaves, J. M., Guda, C., ... Stevens, R. C. (2002). Structural genomics of the Thermotoga maritima proteome implemented in a high-throughput structure determination pipeline. Proceedings of the National Academy of Sciences of the United States of America, 99(18), 11664-11669. https://doi.org/10.1073/pnas.142413399