title: Small Angle X-ray Scattering (SAXS) Characterization of the POTRA Domains of BamA.
abstract: BamA is the central component of the BAM complex and contains a C-terminal β-barrel domain embedded in the outer membrane, and a soluble, periplasmic domain, made out of five polypeptide transport associated (POTRA) motifs. Structural characterization of the POTRA domains was carried out by a combination of crystallographic, NMR and solution Small Angle X-ray Scattering (SAXS) approaches. Despite its limited resolution, SAXS is an excellent complement to NMR and crystallography. It is well suited to validate high-resolution models in solution and is particularly useful to characterize flexible systems such as the POTRA domains of BamA. Here we present a protocol for sample preparation and discuss the considerations of SAXS data collection and quality control, which is applicable to most soluble proteins.
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, CO, 80309, USA.
authors.forename: Pamela Arden
authors.initials: PA
authors.lastname: Doerner
authors.affiliation: Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, CO, 80309, USA. marcelo.sousa@colorado.edu.
authors.forename: Marcelo Carlos
authors.initials: MC
authors.lastname: Sousa
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry*; D029968:Escherichia coli Proteins / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D011786:Quality Control; D053838:Scattering, Small Angle; D012995:Solubility; D014961:X-Ray Diffraction / Q000379:methods*
chemical_list: D001425:Bacterial Outer Membrane Proteins; C503914:BamA protein, E coli; D029968:Escherichia coli Proteins
keywords: BAM; BamA; Conformational change; OMP; SAXS; Structure