Structural Genomics of the Thermotoga maritima Proteome Implemented in a High-Throughput Structure Determination Pipeline

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-throug...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2002-09, Vol.99 (18), p.11664-11669
Main Authors: Lesley, Scott A., Kuhn, Peter, Godzik, Adam, Deacon, Ashley M., Mathews, Irimpan, Kreusch, Andreas, Spraggon, Glen, Klock, Heath E., McMullan, Daniel, Shin, Tanya, Vincent, Juli, Robb, Alyssa, Brinen, Linda S., Miller, Mitchell D., McPhillips, Timothy M., Miller, Mark A., Scheibe, Daniel, Canaves, Jaume M., Guda, Chittibabu, Jaroszewski, Lukasz, Selby, Thomas L., Elsliger, Marc-Andre, Wooley, John, Taylor, Susan S., Hodgson, Keith O., Wilson, Ian A., Schultz, Peter G., Stevens, Raymond C.
Format: Article
Language:English
Subjects:
Bacteria
Biochemistry
Biological Sciences
Cloning, Molecular
Crystal structure
Crystallization
Crystals
Data collection
Genome, Bacterial
Genomics
Models, Molecular
Molecules
Open Reading Frames
Pipelines
Protein Conformation
Proteins
Proteome
Proteomes
Robotics
Thermotoga maritima - genetics
Thermotoga maritima - metabolism
Wave diffraction
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Summary: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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.142413399