Robust, high-throughput solution structural analyses by small angle X-ray scattering (SAXS).
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.
Hura
Greg L
GL
Physical Bioscience Division, Lawrence Berkeley National Laboratory, Berkeley, California, USA.
Menon
Angeli L
AL
Hammel
Michal
M
Rambo
Robert P
RP
Poole
Farris L
FL
Tsutakawa
Susan E
SE
Jenney
Francis E
FE
Classen
Scott
S
Frankel
Kenneth A
KA
Hopkins
Robert C
RC
Yang
Sung-Jae
SJ
Scott
Joseph W
JW
Dillard
Bret D
BD
Adams
Michael W W
MW
Tainer
John A
JA
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*
D001426:Bacterial Proteins; D011506:Proteins