Multi-domain O-GlcNAcase structures reveal allosteric regulatory mechanisms

Hansen S, Bartual S, Yuan H, Raimi O, Gorelik A, Ferenbach A, Lytje K, Pedersen J, Drace T, Boesen T, van Aalten D, Nature Communications 16(1) (2025) DOI

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