Enzymic synthesis of a new type of fucose-containing glycolipid with fucosyltransferase of rat ascites hepatoma cell, AH 7974F.
An alpha-fucosyltransferase activity has been demonstrated in rat ascites hepatoma AH 7974F cells catalyzing the transfer of L-fucose to asialo-GM1 prepared from bovine brain GM1 ganglioside to form a fucolipid in the presence of Triton X-100. The radioactive fucolipid was shown to be Fuc-(alpha1 leads to 2)-Gal-(beta1 leads to 3)-GalNAc-(beta1 leads to 4)-Gal-(beta1 leads to 4)-Glc-ceramide from the following results. The radioactive product coincided with authentic blood group H-active fucolipid from AH 7974F cell on thin-layer chromatography. The product formed a precipitation line not only with Ulex europeus lectin but also with eel anti-H serum on agarose gel plates. The terminal 14C-labeled fucose was released by Bacillus fulminans alpha(1 leads to 2)fucosidase as well as Charonia lampas alpha-fucosidase. The optimum pH value for the incorporation of L-fucose into asialo-GM1 was 5.8 in cacodylate/HCl buffer. The fucosyltransferase was highly specific for asialo-GM1.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002478:Cells, Cultured; D003902:Detergents / Q000494:pharmacology; D005643:Fucose / Q000378:metabolism*; D005647:Fucosyltransferases / Q000378:metabolism*; D005677:G(M1) Ganglioside / Q000378:metabolism; D006028:Glycosphingolipids / Q000096:biosynthesis* / Q000378:metabolism; D006154:Guanosine Diphosphate Fucose / Q000378:metabolism; D006602:Hexosyltransferases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008114:Liver Neoplasms, Experimental / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D051381:Rats
D003902:Detergents; D006028:Glycosphingolipids; D006154:Guanosine Diphosphate Fucose; D005643:Fucose; D005677:G(M1) Ganglioside; D011092:Polyethylene Glycols; D005647:Fucosyltransferases; D006602:Hexosyltransferases; D002118:Calcium