Dictyostelium cytosolic fucosyltransferase synthesizes H type 1 trisaccharide in vitro

A fucosyltransferase activity has been detected using lacto-N-biose I as acceptor in the lower eukaryote Dictyostelium discoideum. This transferase requires divalent cations and is inhibited by N-ethylmaleimide and detergent treatment. Apparent calculated K m values for GDP-Fuc and lacto-N-biose I a...

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Bibliographic Details
Published in:FEBS letters 1996-10, Vol.395 (1), p.68-72
Main Authors: Trinchera, Marco, Bozzaro, Salvatore
Format: Article
Language:English
Subjects:
Acetylglucosamine - analogs & derivatives
Adenosine Triphosphate - pharmacology
Animals
Carbohydrate antigen
Carbohydrate Sequence
Cations, Divalent
Cell Fractionation
Chelating Agents - pharmacology
chloromethyl ketone
Cytosol
Cytosol - enzymology
Dictyostelium - enzymology
Dictyostelium cytosolic α1,2fucosyltransferase
Dictyostelium discoideum
dimethylsulfoxide
DMSO
Dα1,2FucT
Edetic Acid - pharmacology
Enzyme Inhibitors - pharmacology
Ethylmaleimide - pharmacology
Fucose - pharmacology
Fucosyltransferases - analysis
Fucosyltransferases - antagonists & inhibitors
Fucosyltransferases - metabolism
Galactoside 2-alpha-L-fucosyltransferase
Galβ1-4GlcNAc
GDP-Fuc
GDP-β-l-fucose
Glycosyltransferase
Guanosine Diphosphate Fucose - metabolism
H type 1 trisaccharide, Fucα1-2Galβ1-3GlcNAc
high-performance thin-layer chromatography
HPTLC
human blood group H α1,2fucosyltransferase
Hydrogen-Ion Concentration
Hα1,2FucT
Kinetics
LacNAc
lacto-N-biose I, Galβ1-3GlcNAc
Lewis a trisaccharide, Galβ1-3[Fucα1-4]GlcNAc
Magnesium - pharmacology
Manganese - pharmacology
Methylation
Molecular Sequence Data
N-ethylmaleimide
NEM
Permethylation
phenylmethylsulfonyl fluoride
PMSF
Temperature
TLCK
Trisaccharides - biosynthesis
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Description
Summary:A fucosyltransferase activity has been detected using lacto-N-biose I as acceptor in the lower eukaryote Dictyostelium discoideum. This transferase requires divalent cations and is inhibited by N-ethylmaleimide and detergent treatment. Apparent calculated K m values for GDP-Fuc and lacto-N-biose I are 1.27 μM and 2.80 mM, respectively. The activity is quantitatively recovered in the supernatant after centrifugation at 100 000 × g for 1 h. The reaction product, as determined by gel permeation chromatography, sensitivity to fucosidases, and analysis of partially methylated derivatives, is Fucα1-2Galβ1-3GlcNAc (H type 1 trisaccharide).
ISSN:0014-5793
1873-3468
DOI:10.1016/0014-5793(96)01003-4