Unique Structural Features of Mammalian NEIL2 DNA Glycosylase Prime Its Activity for Diverse DNA Substrates and Environments.

Eckenroth BE, Cao VB, Averill AM, Dragon JA, DoubliƩ S, Structure (2020) Europe PMC

SASDJA4 – Endonuclease VIII from E. coli (size exclusion chromatography SAXS)

Endonuclease 8
MWI(0) 29 kDa
MWexpected 31 kDa
VPorod 45 nm3
log I(s) 2.64×102 2.64×101 2.64×100 2.64×10-1
Endonuclease 8 small angle scattering data  s, nm-1
ln I(s)
Endonuclease 8 Guinier plot ln 2.65×102 Rg: 2.3 nm 0 (2.3 nm)-2 s2
Endonuclease 8 Kratky plot 1.104 0 3 sRg
Endonuclease 8 pair distance distribution function Rg: 2.4 nm 0 Dmax: 7.8 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Endonuclease 8 PDB model

log I(s)
 s, nm-1
Endonuclease 8 PDB model

Synchrotron SAXS data from solutions of Endonuclease VIII from E. coli (size exclusion chromatography SAXS) in 25 mM Bis-Tris, 150 mM NaCl, 2% glycerol, 1 mM TCEP, pH 8 were collected on the 12.3.1 (SIBYLS) beam line at the Advanced Light Source (ALS) storage ring (Berkeley, CA, USA) using a Pilatus3 X 2M detector at a wavelength of λ = 0.103 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 70.00 μl sample at 5 mg/ml was injected at a 0.50 ml/min flow rate onto a Shodex KW-800 series column at 25°C. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Endonuclease 8 (EcoNei (R253A))
Mol. type   Protein
Organism   Escherichia coli
Olig. state   Monomer
Mon. MW   30.8 kDa
UniProt   P50465 (2-263)
Sequence   FASTA
PDB code   1Q3B
PDB code   2EA0