High-resolution structure of the vaccinia virus phospholipase D-fold endonuclease K4.

Gröger H Trouba C, Barbaste J, Lacroix G, Schoehn G, Burmeister WP, Tarbouriech N, J Mol Biol :169961 (2026) Europe PMC

SASDZ78 – Octameric Assembly of the Phospholipase D–Like (PLD-Fold) Nuclease K4 Domain in the Vaccinia Virion Nicking-Joining Enzyme

Virion nicking-joining enzyme (octamer)
MWexperimental 407 kDa
MWexpected 393 kDa
VPorod 491 nm3
log I(s) 1.29×10-2 1.29×10-3 1.29×10-4 1.29×10-5
Virion nicking-joining enzyme (octamer) small angle scattering data  s, nm-1
ln I(s)
Virion nicking-joining enzyme (octamer) Guinier plot ln 1.29×10-2 Rg: 5.0 nm 0 (5.0 nm)-2 s2
(sRg)2I(s)/I(0)
Virion nicking-joining enzyme (octamer) Kratky plot 1.104 0 3 sRg
p(r)
Virion nicking-joining enzyme (octamer) pair distance distribution function Rg: 5.1 nm 0 Dmax: 18 nm

Data validation


Fits and models


log I(s)
 s, nm-1
Virion nicking-joining enzyme (octamer) PHENIX model

Synchrotron SAXS data from solutions of Octameric Assembly of the Phospholipase D–Like (PLD-Fold) Nuclease K4 Domain in the Vaccinia Virion Nicking-Joining Enzyme in 20 mM Tris pH 7.5, 150 mM NaCl, 0.5mM TCEP, pH 7.5 were collected on the SWING beam line at the SOLEIL storage ring (Saint-Aubin, France) using a Eiger 4M detector at a sample-detector distance of 2 m and at a wavelength of λ = 0.1033 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 45.00 μl sample at 1 mg/ml was injected at a 0.30 ml/min flow rate onto a GE Superdex 200 Increase 5/150 column at 20°C. 630 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.

Molecular mass calculated from the Porod volume using 0.83 Da∙Å-3 obtained from the protein specific mass of 1,37 g∙cm-3. The sequence of the construct after tag-cleavage is provided.

Virion nicking-joining enzyme (octamer)
Mol. type   Protein
Organism   Vaccinia virus (strain Copenhagen)
Olig. state   Octamer
Mon. MW   49.1 kDa
 
UniProt   P20537 (2-424)
Sequence   FASTA