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SAXS experiments were conducted at the beamline B21 of the Diamond Light Source (Didcot, UK). CCDC6-RET∆CT (40 µL, 5 mg/mL) was injected at room temperature into a Superdex 200 Increase 3.2 column connected to an in-line Agilent 1200 HPLC system. The buffer used for the SEC column was the same protein buffer (20 mM Tris pH 7.5, 500 mM NaCl, 1 mM DTT). The eluted samples were exposed for 3s and 600 frames taken using an X-ray wavelength of 1 nm, and a sample to detector (Eiger 4M) distance of 3.7 m. The data covered a momentum transfer range of 0.032 < s < 3.4 nm-1. The frames recorded immediately before elution of the sample were subtracted from the protein scattering profiles. The Scåtter software package was used to analyze data, buffer-subtraction, scaling, merging and checking possible radiation damage of the samples.
The Rg value was calculated with the Guinier approximation assuming that at very small angles s < 1.3/Rg. The particle distance distribution, Dmax, was calculated from the scattering pattern with GNOM, and shape estimation was carried out with DAMMIF/DAMMIN, all these programs included in the ATSAS package.
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s, nm-1
