Synchrotron SAXS
data from solutions of
A nuclear mRNA-binding protein bound to GT-repeat DNA Oligo: TDP-43 (TAR DNA-binding protein 43)
in
20 mM HEPES, 100 mM KCl, 2 mM TCEP, pH 7.6
were collected
on the
SWING beam line
at the SOLEIL storage ring
(Saint-Aubin, France)
using a Eiger 4M detector
at a sample-detector distance of 2 m and
at a wavelength of λ = 0.1033 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 1 mg/ml was injected at a 0.30 ml/min flow rate
onto a Agilent AdvanceBio SEC 300Å, 4.6 x 150 mm column
at 25°C.
600 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.
15 µL of ssDNA (GT)12 was incubated with 50 µL of TDP-43 during 1 hour at room temperature to reach a final ratio for the complex TDP-43:ssDNA of 2:1 with 60 µM of TDP-43 and 27 µM of ssDNA. 50 µL of this complex was injected into the SEC column.
s, nm-1
