Synchrotron SAXS
data from solutions of
Fep1-GATA
in
50 mM MOPS, 50 mM NaCl, pH 7
were collected
on the
BM29 beam line
at the ESRF storage ring
(Grenoble, France)
using a Pilatus 1M detector
at a sample-detector distance of 2.9 m and
at a wavelength of λ = 0.0999 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 40.00 μl sample
was injected at a 0.40 ml/min flow rate
onto a GE Superdex 75 Increase 5/150 column
at 20°C.
900 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 complex between the Fep1 transcription factor and its cognate DNA (GATAds) has been formed in solution prior to injection to the HPLC-SEC column. Protein and DNA were both 100 microM, the expected MW for a 1:1 complex is 34 kDa.
The experiment has been run at BM29 of ESRF (Grenoble, France) on the same day as the previous deposited samples. All the protein samples were coming from the same purification batch. Data were analysed with ChromixS, for buffer subtraction 20 frames were averaged, the I(q) vs q was derived form the frames of the main peak (256-282).
s, nm-1
