Multi-domain O-GlcNAcase structures reveal allosteric regulatory mechanisms

Jan Skov Pedersen.

SASDXF6 – Multi-domain human O-GlcNAcase

Protein O-GlcNAcase
MWexperimental 278 kDa
MWexpected 163 kDa
VPorod 443 nm3
log I(s) 5.27×10-1 5.27×10-2 5.27×10-3 5.27×10-4
Protein O-GlcNAcase small angle scattering data  s, nm-1
ln I(s)
Protein O-GlcNAcase Guinier plot ln 5.27×10-1 Rg: 5.4 nm 0 (5.4 nm)-2 s2
(sRg)2I(s)/I(0)
Protein O-GlcNAcase Kratky plot 1.104 0 √3 sRg
p(r)
Protein O-GlcNAcase pair distance distribution function Rg: 5.5 nm 0 Dmax: 18.9 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Protein O-GlcNAcase ALPHAFOLD model
SAXS data from solutions of Multi-domain human O-GlcNAcase in 25 mM Tris buffer, 150 mM NaCl, and 0.5 mM TCEP, pH 7.5 were collected on the Bruker Nanostar w Excillum source instrument (Department of Chemistry, iNANO building, Aarhus Uinversity, Aarhus C, Denmark) using a VÅNTEC-2000 detector at a sample-detector distance of 0.9 m and at a wavelength of λ = 1.34 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). One solute concentration of 2.50 mg/ml was measured at 20°C. One 1800 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.

Protein O-GlcNAcase (h-OGA)
Mol. type   Protein
Organism   Homo sapiens
Olig. state   Dimer
Mon. MW   81.5 kDa
 
UniProt   O60502 (11-766)
Sequence   FASTA