ENZYMIC-SYNTHESIS OF A NEW TYPE OF FUCOSE-CONTAINING GLYCOLIPID WITH FUCOSYLTRANSFERASE OF RAT ASCITES HEPATOMA CELL, AH-7974F

  • 1 January 1979
    • journal article
    • research article
    • Vol. 572  (1) , 105-112
Abstract
An .alpha.-fucosyltransferase activity was demonstrated in rat ascites hepatoma AH 7974F cells catalyzing the transfer of L-fucose to asialo-GM1 [Gal-(.beta.1 .fwdarw. 3)-GalNAc-(.beta.l .fwdarw. 4)-Gal-(.beta.l-4)-Glc-ceramide] prepared from bovine brain GM1 ganglioside to form a fucolipid in the presence of Triton X-100. The radioactive fucolipid was Fuc(.alpha.1 .fwdarw. 2)asialo-GM1. The radioactive product coincided with authentic blood group H-active fucolipid from AH 7974F cell on TLC. The product formed a precipitation line with Ulex europaeus lectin and with eel anti-H serum on agarose gel plates. The terminal 14C-labeled fucose was released by Bacillus fulminans .alpha.(1 .fwdarw. 2)fucosidase and by 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.