|
Synchrotron SAXS data from solutions of cone-rod homeobox protein (a peptide of the DNA binding domain of CRX) in 50 mM sodium phosphate,100 mM NaCl, and 5 mM imidazole, pH 7 were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS; Berkeley, CA, USA) using a Pilatus3 X 2M detector at a sample-detector distance of 1.5 m and at a wavelength of λ = 0.103 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). This setup results in scattering vectors, s, ranging from 0.13 nm-1 to 5 nm-1. In-line size-exclusion chromatography (SEC) SAS was employed. The SEC parameters were as follows: A sample at 7 mg/ml was injected at a 0.50 ml/min flow rate onto a Shodex KW-802.5 analytical column at 10°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.
The CRX DBD peptide was resuspended in water before being dialyzed overnight at 4 ˚C into 50 mM sodium phosphate, pH 7, 100 mM NaCl, and 5 mM imidazole. Samples were shipped overnight at 4 °C to the SIBYLS beamline at the Advanced Light Source.
|
|
s, nm-1
