Oligomeric State of β-Coronavirus Non-Structural Protein 10 Stimulators Studied by Small Angle X-ray Scattering

Knecht W, Fisher S, Lou J, Sele C, Ma S, Rasmussen A, Pinotsis N, Kozielski F, International Journal of Molecular Sciences 24(17):13649 (2023) DOI

SASDS89 – Short SARS-CoV-2 Non-structural protein 10 (Nsp10: N- and C-terminal truncation variant)

Replicase polyprotein 1ab (Non-structural protein 10 - Δ4254-4262; Δ4386-4392)
MWI(0) 9 kDa
MWexpected 13 kDa
VPorod 16 nm3
log I(s) 7.92×10-3 7.92×10-4 7.92×10-5 7.92×10-6
Replicase polyprotein 1ab (Non-structural protein 10 - Δ4254-4262; Δ4386-4392) small angle scattering data  s, nm-1
ln I(s)
Replicase polyprotein 1ab (Non-structural protein 10 - Δ4254-4262; Δ4386-4392) Guinier plot ln 7.92×10-3 Rg: 1.5 nm 0 (1.5 nm)-2 s2
(sRg)2I(s)/I(0)
Replicase polyprotein 1ab (Non-structural protein 10 - Δ4254-4262; Δ4386-4392) Kratky plot 1.104 0 3 sRg
p(r)
Replicase polyprotein 1ab (Non-structural protein 10 - Δ4254-4262; Δ4386-4392) pair distance distribution function Rg: 1.6 nm 0 Dmax: 5.1 nm

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of Short SARS-CoV-2 Non-structural protein 10 in 10 mM HEPES, 300 mM NaCl, 1.5% (v/v) glycerol, 2 mM DTT, pH 7.5 were collected on the B21 beam line at the Diamond Light Source (Didcot, UK) using a Eiger 4M detector at a sample-detector distance of 3.7 m and at a wavelength of λ = 0.09464 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 35.00 μl sample at 8 mg/ml was injected at a 0.08 ml/min flow rate onto a Cytiva Superdex 75 10/300 column at 4°C. 21 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.

Non-structural protein 10 from SARS-CoV-2 with N- and C-terminal amino acid truncations.

Replicase polyprotein 1ab (Non-structural protein 10 - Δ4254-4262; Δ4386-4392) (Nsp10)
Mol. type   Protein
Organism   Severe acute respiratory syndrome coronavirus 2 (2019-nCoV) (SARS-CoV-2)
Olig. state   Monomer
Mon. MW   13.3 kDa
 
UniProt   P0DTD1 (4263-4385)
Sequence   FASTA