Synchrotron SAXS data from solutions of the Aedes aegypti Toll5A ectodomain in complex with Spz1C Cys-knot domain in 50 mM NaCl, 50 mM Tris-HCl, pH 7.5 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Eiger 4M detector at a sample-detector distance of 3.7 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 55.00 μl sample at 5 mg/ml was injected at a 0.075 ml/min flow rate onto a GE Superdex 6 Increase 3.2/300 column at 20°C. 31 successive 3 second frames were collected through the SEC elution peak of the sample. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.
The Toll5A-Spz1C complex was concentrated to 5 mg/mL in 50mM NaCl, 50 mM Tris-HCl pH7.5 and injected into a Superose 6 Increase 3.2 equilibrated in the same buffer at a flow rate of 0.075 ml/min. Data points were collected every 3 sec.