Keil P,
Wulf A,
Kachariya N,
Reuscher S,
Hühn K,
Silbern I,
Altmüller J,
Keller M,
Stehle R,
Zarnack K,
Sattler M,
Urlaub H,
Sträßer K,
Nucleic Acids Res
(2022)
Europe PMC
SASDQV5 – NPL3 protein RNA recognition motifs 1 and 2 (RRM1,2) wild-type, bound to RNA
SAXS data from solutions of the NPL3 protein RNA recognition motif domains 1 and 2 (RRM1,2) bound to RNA in 20 mM NaPO4, 50 mM NaCl, 1 mM DTT, pH 6.5 were collected using a Rigaku bioSAXS-1000 instrument at the Technische Universität München (TUM; Garching, Germany) equipped with a Pilatus 100K detector at a sample-detector distance of 0.5 m and at a wavelength of λ = 0.155 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 4.90 mg/ml was measured at 25°C. Eight successive 900 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 RNA bound NPL3 complex was prepared by mixing the equimolar ratio of protein to RNA. The complex was further purified using gel-filtration chromatography. The short RNA sequence 5'-AGCACCGUGGAGA-3' was used in preparation of protein-RNA complex.