Integrative SAXS and AFM analysis of engineered carbohydrate-active enzyme assemblies with tunable spatial organization.

Pardo Larrabeiti I, Eibinger M, Esque J, Pradeau S, Fort S, Moraïs S, Mizrahi I, Bayer EA, Nidetzky B, Montanier CY, Roblin P, Dumon C, Protein Sci 35(7):e70649 (2026) Europe PMC

SASDYT7 – Glycoside Hydrolase family 11 (AtXyn11A) from Acetovibrio thermocellus

Endoxylanase Xyn11A
MWexperimental 40 kDa
MWexpected 40 kDa
VPorod 50 nm3
log I(s) 3.47×10-3 3.47×10-4 3.47×10-5 3.47×10-6
Endoxylanase Xyn11A small angle scattering data  s, nm-1
ln I(s)
Endoxylanase Xyn11A Guinier plot ln 3.47×10-3 Rg: 2.9 nm 0 (2.9 nm)-2 s2
(sRg)2I(s)/I(0)
Endoxylanase Xyn11A Kratky plot 1.104 0 √3 sRg
p(r)
Endoxylanase Xyn11A pair distance distribution function Rg: 3.1 nm 0 Dmax: 10 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Endoxylanase Xyn11A GASBOR model
SAXS data from solutions of Glycoside Hydrolase family 11 (AtXyn11A) from Acetovibrio thermocellus in 25 mM Tris, 150 mM NaCl, pH 8 were collected on the XEUSS 2.0 instrument (Laboratoire de Génie Chimique, Toulouse, France) using a Pilatus 1M detector (DECTRIS, Switzerland) detector at a sample-detector distance of 1.3 m and at a wavelength of λ = 0.154 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 3 and 36 mg/ml were measured at 20°C. 12 successive 600 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. The low angle data collected at lower concentration were merged with the highest concentration high angle data to yield the final composite scattering curve.

Endoxylanase Xyn11A (Xyn11A)
Mol. type   Protein
Organism   Acetovibrio thermocellus
Olig. state   Monomer
Mon. MW   39.8 kDa
 
UniProt   O52779
Sequence   FASTA