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Synchrotron SAXS
data from solutions of
Fep1 wild type with [2Fe2S] cluster reconstituted
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
at 3.4 mg/ml 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.
Purified Fep1 was reconstituted with 2 stoichiometric excess of (FeCl3 and Na2S) in 50 mM MOPS, 50 mM NaCl, 1 mM TCEP pH 7.0, then the excess of Fe was eliminated by passing the solution into a PD10 column.
Then the purified reconstituted protein was concentrated and the concentration was measured to be 3.43 mg/ml.
Two successive HPLC run were performed on two injections by 40 microL each and the frames from the 2 main elution peaks were merged and analysed with Chromixs. The I(q) vs q were solvent subtracted by using the first 20 frames of the HPLC run, after being averaged.
The same SEC column as for Fep1 wild type was used.
Data were collected at BM29 of ESRF (Grenoble, France)
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s, nm-1
![Fep1 wild type with [2Fe2S] cluster reconstituted Rg histogram](/media/fitting_files/extra/plots/SASDJ76_fit1_extra1_rghistogram_img.png "Fep1 wild type with [2Fe2S] cluster reconstituted Rg histogram")
Rg, nm
