Synchrotron SAXS data from FhuF in 20 mM Phosphate, 200 mM NaCl, pH 7.4 was collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Pilatus 2M detector at a sample-detector distance of 2.7 m and at a wavelength of λ = 0.091 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle).
In-line size-exclusion chromatography (SEC) SAS was employed using an Agilent 1200 HPLC System by injecting a 50.00 μl sample at 8.9 mg/ml under a flow rate of 0.16 ml/min onto a Shodex KW402.5-4F column (Showa Denko) at 10°C collecting 915 successive 2-second frames.
The data were normalized to the intensity of the transmitted beam and radially averaged; the solvent-blank scattering was subtracted from the sample elution’s peak frames.
The displayed SAXS-generated ab initio molecular reconstruction was obtained using DAMMIF (DAMFILT occupancy and volume-corrected bead model) by clustering and averaging dummy residue models from 20 independent runs, with an averaged Normalized Spatial Discrepancy (NSD) of 0.727.
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