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Synchrotron SAXS data from a solution of Men2 ACP1ACP2TE protein in 25 mM Tris 150 mM NaCl, pH 8, was collected using in-line SEC-SAXS at the B21 beam line at the Diamond Light Source (Oxfordshire, UK) using a Pilatus 2M detector at a sample-detector distance of 4.0 m and at a wavelength of λ = 0.1 nm (l(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). In line SEC-SAXS was performed using an Agilent 1200 HPLC system equipped with a 2.4 mL Superdex S200 column (3.2/300 GE Healthcare). A sample concentration of 6.9 mg/ml (45 μl) was loaded onto the column at a flow rate of 0.075 mL/min. Data frames were collected at 25°C for the entire column elution with an exposure time of 3 seconds per frame. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted using data frames measured from the flow-through column buffer with no macromolecular sample present.
The model fits, Rg distributions and a sub-set of ensemble model representatives as shown describe: Top: EOM of Men2 ACP1ACP2TE when modelling from 2 pools of apo and holo Men2 ACP1ACP2TE whereby ratios of 46% of apo Men2 ACP1ACP2TE and 54% of holo Men2 ACP1ACP2TE were applied to model a mixture of the two forms. Middle: EOM of Men2 ACP1ACP2TE when modelling as apo Men2 ACP1ACP2TE. Bottom: EOM of Men2 ACP1ACP2TE when modelling as holo Men2 ACP1ACP2TE. For clarity, only 3 ensemble model representatives from the combined pool are shown. The complete EOM-refined model-pool representatives and EOM log files are made available in the full entry zip archive.
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s, nm-1
Rg, nm
Rg, nm
Rg, nm
