title: Robust, high-throughput solution structural analyses by small angle X-ray scattering (SAXS).
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.
authors.affiliation: Physical Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
authors.forename: Greg L
authors.initials: GL
authors.lastname: Hura
authors.forename: Angeli L
authors.initials: AL
authors.lastname: Menon
authors.forename: Michal
authors.initials: M
authors.lastname: Hammel
authors.forename: Robert P
authors.initials: RP
authors.lastname: Rambo
authors.forename: Farris L
authors.initials: FL
authors.lastname: Poole
authors.forename: Susan E
authors.initials: SE
authors.lastname: Tsutakawa
authors.forename: Francis E
authors.initials: FE
authors.lastname: Jenney
authors.forename: Scott
authors.initials: S
authors.lastname: Classen
authors.forename: Kenneth A
authors.initials: KA
authors.lastname: Frankel
authors.forename: Robert C
authors.initials: RC
authors.lastname: Hopkins
authors.forename: Sung-Jae
authors.initials: SJ
authors.lastname: Yang
authors.forename: Joseph W
authors.initials: JW
authors.lastname: Scott
authors.forename: Bret D
authors.initials: BD
authors.lastname: Dillard
authors.forename: Michael W W
authors.initials: MW
authors.lastname: Adams
authors.forename: John A
authors.initials: JA
authors.lastname: Tainer
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry; D004867:Equipment Design; D008958:Models, Molecular; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D020027:Pyrococcus furiosus / Q000378:metabolism; D053838:Scattering, Small Angle; D014961:X-Ray Diffraction / Q000295:instrumentation / Q000379:methods*
chemical_list: D001426:Bacterial Proteins; D011506:Proteins