title: The application of hybrid pixel detectors for in-house SAXS instrumentation with a view to combined chromatographic operation.
abstract: This study analyses the potential for laboratory-based size-exclusion chromatography (SEC) integrated small-angle X-ray scattering (SAXS) instrumentation to characterize protein complexes. Using a high-brilliance home source in conjunction with a hybrid pixel X-ray detector, the efficacy of SAXS data collection at pertinent protein concentrations and exposure times has been assessed. Scattering data from SOD1 and from the complex of SOD1 with its copper chaperone, using 10 min exposures, provided data quality in the range 0.03 < q < 0.25 Å(-1) that was sufficient to accurately assign radius of gyration, maximum dimension and molecular mass. These data demonstrate that a home source with integrated SEC-SAXS technology is feasible and would enable structural biologists studying systems containing transient protein complexes, or proteins prone to aggregation, to make advanced preparations in-house for more effective use of limited synchrotron beam time.
authors: Molecular Biophysics Group, Institute of Integrative Biology, Faculty of Health and Life Sciences, University of Liverpool, Liverpool, UK.
authors: Gareth S A
authors: GS
authors: Wright
authors: Hyun Chul
authors: HC
authors: Lee
authors: Clemens
authors: C
authors: Schulze-Briese
authors: J Günter
authors: JG
authors: Grossmann
authors: Richard W
authors: RW
authors: Strange
authors: S Samar
authors: SS
authors: Hasnain
mesh_terms: D002850:Chromatography, Gel / Q000295:instrumentation*; D006801:Humans; D018832:Molecular Chaperones / Q000737:chemistry; D011487:Protein Conformation; D020836:Protein Structure, Quaternary; D011506:Proteins / Q000737:chemistry*; D053838:Scattering, Small Angle; D013482:Superoxide Dismutase / Q000737:chemistry; D000072105:Superoxide Dismutase-1; D014961:X-Ray Diffraction / Q000295:instrumentation* / Q000379:methods; D014965:X-Rays
chemical_list: C114678:CCS protein, human; D018832:Molecular Chaperones; D011506:Proteins; C000606290:SOD1 protein, human; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1
keywords: SAXS; chromatography; home source; hybrid pixel X-ray detector; protein–protein complex