Synchrotron SAXS data from solutions of Bamb_5924 NDD mutant K32A in 20 mM Tris-HCl, 200 mM NaCl, 5% glycerol, pH 7.5 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a AVIEX PCCD170170 detector at a sample-detector distance of 1.8 m and at a wavelength of λ = 0.1 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A 50.00 μl sample at 28 mg/ml was injected at a 0.10 ml/min flow rate onto a Agilent Bio SEC-3, 100 Å column at 15°C. 100 successive 0.750 second frames were collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.
The protein samples were injected using the online automatic sample changer into a pre-equilibrated HPLC-coupled size-exclusion chromatography column.