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