The first structure–function study of GH151 α‐ l ‐fucosidase uncovers new oligomerization pattern, active site complementation, and selective substrate specificity

Koval'ová T, Kovaľ T, Stránský J, Kolenko P, Dušková J, Švecová L, Vodičková P, Spiwok V, Benešová E, Lipovová P, Dohnálek J, The FEBS Journal (2022) DOI

SASDKY7 – α-L-Fucosidase isoenzyme 2 from Paenibacillus thiaminolyticus - wild type

Alpha-L-fucosidase
MWexperimental 318 kDa
MWexpected 297 kDa
VPorod 418 nm3
log I(s) 2.56×101 2.56×100 2.56×10-1 2.56×10-2
Alpha-L-fucosidase small angle scattering data  s, nm-1
ln I(s)
Alpha-L-fucosidase Guinier plot ln 2.56×101 Rg: 4.2 nm 0 (4.2 nm)-2 s2
(sRg)2I(s)/I(0)
Alpha-L-fucosidase Kratky plot 1.104 0 √3 sRg
p(r)
Alpha-L-fucosidase pair distance distribution function Rg: 4.6 nm 0 Dmax: 13.2 nm

Data validation

Fits and models

log I(s)
 s, nm-1
Alpha-L-fucosidase OTHER model
log I(s)
 s, nm-1
Alpha-L-fucosidase DAMMIN model
SAXS data from solutions of wild-type α-L-Fucosidase isoenzyme 2 in 50 mM potassium phosphate, pH 7.4 were collected using an Anton Paar SAXSpoint 2.0 instrument (Institute of Biotechnology, Czech Academy of Sciences, Vestec, Czech Republic) equipped with an Eiger R 1M detector at a sample-detector distance of 0.8 m and at a wavelength of λ = 0.134 nm (I(s) vs s, where s = 4πsinθ/λ, and 2θ is the scattering angle). Solute concentrations ranging between 0.8 and 5.7 mg/ml were measured at 15°C. 15 successive 60 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.

Storage temperature = UNKNOWN

Alpha-L-fucosidase (Fuk2 wt)
Mol. type   Protein
Organism   Paenibacillus thiaminolyticus (Bacillus thiaminolyticus)
Olig. state   Tetramer
Mon. MW   74.3 kDa
 
UniProt   K0JCW6 (1-660)
Sequence   FASTA