Herpes simplex virus 1 protein pUL21 alters ceramide metabolism by activating the inter-organelle transport protein CERT

Benedyk T, Connor V, Caroe E, Shamin M, Svergun D, Deane J, Jeffries C, Crump C, Graham S, Journal of Biological Chemistry :102589 (2022) DOI

SASDNB7 – C-terminal domain of pUL21 with N-terminal His-tag

Tegument protein UL21 (C-terminal domain)
MWexperimental 32 kDa
MWexpected 29 kDa
VPorod 49 nm3
log I(s) 1.23×10-2 1.23×10-3 1.23×10-4 1.23×10-5
Tegument protein UL21 (C-terminal domain) small angle scattering data  s, nm-1
ln I(s)
Tegument protein UL21 (C-terminal domain) Guinier plot ln 1.24×10-2 Rg: 2.2 nm 0 (2.2 nm)-2 s2
(sRg)2I(s)/I(0)
Tegument protein UL21 (C-terminal domain) Kratky plot 1.104 0 √3 sRg
p(r)
Tegument protein UL21 (C-terminal domain) pair distance distribution function Rg: 2.2 nm 0 Dmax: 8.5 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Tegument protein UL21 (C-terminal domain) GASBOR model
log I(s)
 s, nm-1
Tegument protein UL21 (C-terminal domain) CORAL model
log I(s)
 s, nm-1
Tegument protein UL21 (C-terminal domain) CORAL model
Synchrotron SAXS data from solutions of C-terminal domain of pUL21 with N-terminal His-tag in 20 mM Tris pH 8.5, 500 mM NaCl, 3% (v/v) glycerol, 1 mM DTT, pH 8.5 were collected on the EMBL P12 beam line at the PETRA III storage ring (DESY; Hamburg, Germany) using a Pilatus 6M detector at a sample-detector distance of 3 m and at a wavelength of λ = 0.124 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 0.68 mg/ml was measured at 20°C. 47 successive 0.100 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.

Tegument protein UL21 (C-terminal domain) (H6-pUL21C)
Mol. type   Protein
Organism   Human herpesvirus 1 (strain 17)
Olig. state   Monomer
Mon. MW   29.3 kDa
 
UniProt   P10205 (275-535)
Sequence   FASTA