1. Robust, high-throughput solution structural analyses by small angle X-ray scattering (SAXS).
- Author
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Hura GL, Menon AL, Hammel M, Rambo RP, Poole FL 2nd, Tsutakawa SE, Jenney FE Jr, Classen S, Frankel KA, Hopkins RC, Yang SJ, Scott JW, Dillard BD, Adams MW, and Tainer JA
- Subjects
- Bacterial Proteins chemistry, Equipment Design, Models, Molecular, Protein Conformation, Pyrococcus furiosus metabolism, X-Ray Diffraction instrumentation, Proteins chemistry, Scattering, Small Angle, X-Ray Diffraction methods
- Abstract
We present an efficient pipeline enabling high-throughput analysis of protein structure in solution with small angle X-ray scattering (SAXS). Our SAXS pipeline combines automated sample handling of microliter volumes, temperature and anaerobic control, rapid data collection and data analysis, and couples structural analysis with automated archiving. We subjected 50 representative proteins, mostly from Pyrococcus furiosus, to this pipeline and found that 30 were multimeric structures in solution. SAXS analysis allowed us to distinguish aggregated and unfolded proteins, define global structural parameters and oligomeric states for most samples, identify shapes and similar structures for 25 unknown structures, and determine envelopes for 41 proteins. We believe that high-throughput SAXS is an enabling technology that may change the way that structural genomics research is done.
- Published
- 2009
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