| MWexperimental | 91 | kDa |
| MWexpected | 84 | kDa |
| VPorod | 183 | nm3 |
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log I(s)
5.17×10-2
5.17×10-3
5.17×10-4
5.17×10-5
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s, nm-1
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Alternative dimerization is required for activity and inhibition of the HEPN ribonuclease RnlA
Garcia-Rodriguez G, Charlier D, Wilmaerts D, Michiels J, Loris R, Nucleic Acids Research 49(12):7164-7178 (2021) DOISASDKS9 – mRNA endoribonuclease toxin LS (D245R mutant); batch-SAXS measurements in low salt buffer
mRNA endoribonuclease toxin LS (D245R mutant)|
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Fits and models
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Synchrotron SAXS data from solutions of D245R RnlA in 20 mM Tris, 150 mM NaCl, 1 mM TCEP, pH 8 were collected on the SWING beam line at SOLEIL (Saint-Aubin, France) using a Eiger 4M detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.10331 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.3 and 2.2 mg/ml were measured at 16°C. 10 successive 0.990 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 low angle data collected at lower concentrations were extrapolated to infinite dilution and merged with the higher concentration data to yield the final composite scattering curve.
A monodisperse solution of D245R RnlA protein at 3 mg/ml was serially diluted to 2.2, 0.89, 0.66, and 0.34 mg/ml. This concentration series was measured in batch mode and curves were buffer subtracted and averaged. Infinite dilution extrapolation was performed by merging the scattering intensity of the most diluted concentration point at the lowest q values with the averaged curve. |
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