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 Km values for GDP-Fuc and lacto-N-biose I are 1.27 microM and 2.80 mM, respectively. The activity is quantitatively recovered in the supernatant after centrifugation at 100000 x g for 1 h. The reaction product, as determined by gel permeation chromatography, sensitivity to fucosidases, and analysis of partially methylated derivatives, is Fucalpha1-2Galbeta1-3GlcNAc (H type 1 trisaccharide).
Trinchera
M
M
Department of Biochemistry, University of Pavia, Italy.
Bozzaro
S
S
D000117:Acetylglucosamine / Q000031:analogs & derivatives; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002240:Carbohydrate Sequence; D002413:Cations, Divalent; D002458:Cell Fractionation; D002614:Chelating Agents / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology*; D004023:Dictyostelium / Q000201:enzymology*; D004492:Edetic Acid / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005033:Ethylmaleimide / Q000494:pharmacology; D005643:Fucose / Q000494:pharmacology; D005647:Fucosyltransferases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006154:Guanosine Diphosphate Fucose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008745:Methylation; D008969:Molecular Sequence Data; D013696:Temperature; D014312:Trisaccharides / Q000096:biosynthesis*; D000097631:Galactoside 2-alpha-L-fucosyltransferase
D000117:Acetylglucosamine; D000255:Adenosine Triphosphate; D002413:Cations, Divalent; D002614:Chelating Agents; D004492:Edetic Acid; D004791:Enzyme Inhibitors; D005033:Ethylmaleimide; D005643:Fucose; D005647:Fucosyltransferases; D006154:Guanosine Diphosphate Fucose; D008274:Magnesium; D008345:Manganese; D014312:Trisaccharides; D000097631:Galactoside 2-alpha-L-fucosyltransferase; C033398:galactosyl-1,3-N-acetylglucosamine