SAXS studies of X-ray induced disulfide bond damage: Engineering high-resolution insight from a low-resolution technique

Stachowski T, Snell M, Snell E, Boggon T, PLOS ONE 15(11):e0239702 (2020) DOI

SASDHF7 – N-terminal engineered, disulfide-containing endo-beta-N-acetylglucosaminidase H at 1161.6 Gy X-ray dose (J/kg)

Endo-beta-N-acetylglucosaminidase H
MWexperimental 39 kDa
MWexpected 61 kDa
VPorod 61 nm3
log I(s) 1.72×101 1.72×100 1.72×10-1 1.72×10-2
Endo-beta-N-acetylglucosaminidase H small angle scattering data  s, nm-1
ln I(s)
Endo-beta-N-acetylglucosaminidase H Guinier plot ln 1.72×101 Rg: 2.8 nm 0 (2.8 nm)-2 s2
(sRg)2I(s)/I(0)
Endo-beta-N-acetylglucosaminidase H Kratky plot 1.104 0 3 sRg

Data validation


There are no models related to this curve.

Synchrotron SAXS data from solutions of N-terminal engineered, disulfide-containing endo-beta-N-acetylglucosaminidase H at 1161.6 Gy X-ray dose (J/kg) in 20 mM Tris-HCl, 50 mM NaCl, 5 mM EDTA, pH 7.5 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 sample-detector distance of 2 m and at a wavelength of λ = 0.1127 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 5.00 mg/ml was measured at 10°C. One 0.300 second frame was collected. The data were normalized to the intensity of the transmitted beam and radially averaged; the scattering of the solvent-blank was subtracted.

Molecular weight was estimated from the volume of correlation (Vc; Rambo & Tainer 2013). Dose was estimated using RADDOSE modified for SAXS (Brooks-Barlett et al., 2017).

Endo-beta-N-acetylglucosaminidase H (Endo H)
Mol. type   Protein
Organism   Streptomyces plicatus
Olig. state   Dimer
Mon. MW   30.4 kDa
 
UniProt   P04067 (47-313)
Sequence   FASTA