Synchrotron SAXS
data from solutions of
Karrikin insensitive 2, KAI2 protein mutant S95A with a synthetic strigolactone analogue ligand (aggregate peak)
in
20 mM HEPES, 150 mM imidazole, 150 mM NaCl, 10% (v/v) glycerol, pH 7.5
were collected
on the
SAXS/WAXS beam line
at the Australian Synchrotron storage ring
(Melbourne, Australia)
using a Pilatus3 S 2M detector
at a sample-detector distance of 2.5 m and
at a wavelength of λ = 0.10332 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 50.00 μl sample
at 8.1 mg/ml was injected at a 0.40 ml/min flow rate
onto a GE Superdex 200 Increase 5/150 column
at 24°C.
666 successive
1 second frames were collected.
The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.
The S95A KAI2 mutant protein was incubated with the synthetic strigolactone analogue desmethyl-GR24-enantiomer-5DS (dGR24ent-5DS). The experimental MW value was determined using the volume of correlation, Vc, method.
s, nm-1
