Co-purification of the Lewis blood group N-acetylglucosaminide alpha 1 goes to 4 fucosyltransferase and an N-acetylglucosaminide alpha 1 goes to 3 fucosyltransferase from human milk.
The Lewis blood group-specified N-acetylglucosaminide alpha 1 goes to 4 fucosyltransferase and an N-acetylglucosaminide alpha 1 goes to 3 fucosyltransferase have been copurified over 500,000-fold from human milk by affinity chromatography on GDP-hexanolamine agarose. The purified enzyme preparation migrates as two major bands on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with apparent Mr = 53,000 and 51,000. Analysis of the acceptor substrate specificity of the transferase(s) and structural characterization of the reaction products indicate that the enzyme(s) forms the Fuc alpha 1 goes to 4GlcNAc, Fuc alpha 1 goes to 3GlcNAc, and Fuc alpha 1 goes to 3Glc linkages with oligosaccharide acceptors containing the nonreducing terminal sequences Gal beta 1 goes to 3GlcNAc, Gal beta 1 goes to 4GlcNAc, and Gal beta 1 goes to 4Glc, respectively. The two fucosyltransferase activities are activated to the same extent by a variety of divalent metal ions, inactivated at identical rates by thermal denaturation or reaction with N-ethylmaleimide, and inhibited to the same extent by rabbit antiserum prepared against the purified fucosyltransferase(s). In addition, kinetic analysis of the initial rate data obtained using acceptors for one of the fucosyltransferase activities as an inhibitor of the second suggests that acceptors for both fucosyltransferase activities bind at a single active site.
D000936:Antigen-Antibody Complex; D002413:Cations, Divalent; D005260:Female; D005647:Fucosyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D006602:Hexosyltransferases / Q000302:isolation & purification*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007106:Immune Sera; D007700:Kinetics; D007983:Lewis Blood Group Antigens; D008895:Milk, Human / Q000201:enzymology*; D008970:Molecular Weight; D011247:Pregnancy; D013379:Substrate Specificity
D000936:Antigen-Antibody Complex; D002413:Cations, Divalent; D007106:Immune Sera; D007983:Lewis Blood Group Antigens; D005647:Fucosyltransferases; D006602:Hexosyltransferases; C031692:galactoside 3-fucosyltransferase; C019818:3-galactosyl-N-acetylglucosaminide 4-alpha-L-fucosyltransferase