Synchrotron SAXS data from solutions of Aquifex aeolicus McoA metaloxidase evolved variant (2F4) in 50 mM Tris-HCl, 150 mM NaCl, 2 mM TCEP, pH 7.5 were collected on the BM29 beam line at the ESRF (Grenoble, France) using a Pilatus 1M detector at a sample-detector distance of 2.9 m and at a wavelength of λ = 0.099 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 250 μl sample at 10 mg/ml was injected at a 0.50 ml/min flow rate onto a GE Superdex 200 Increase 10/300 column at 20°C. 3000 successive 1 second frames were collected through the SEC elution. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted from the sample peak data frames.
The scattering intensity from the monomeric elution single-peak region were integrated and buffer subtracted to produce the SAXS-profile of 2F4 using the ScÅtter software. Further processing was performed using the ATSAS software suite.
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