Robust, high-throughput solution structural analyses by small angle X-ray scattering (SAXS)

Nat Methods. 2009 Aug;6(8):606-12. doi: 10.1038/nmeth.1353. Epub 2009 Jul 20.

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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacterial Proteins / chemistry
  • Equipment Design
  • Models, Molecular
  • Protein Conformation
  • Proteins / chemistry*
  • Pyrococcus furiosus / metabolism
  • Scattering, Small Angle*
  • X-Ray Diffraction / instrumentation
  • X-Ray Diffraction / methods*

Substances

  • Bacterial Proteins
  • Proteins