Use of N-acetyl-2'-O-methyllactosamine as a specific acceptor for the determination of alpha-L-(1----3)-fucosyltransferase in human serum.
A synthetic substrate, N-acetyl-2'-O-methyllactosamine, was employed as a specific acceptor for alpha-L-(1----3)-fucosyltransferase from human serum. The fucosyl linkage of the product from this substrate was characterized by hydrolysis with a specific alpha-L-(1----3)/(1----4)-fucosidase. Using this acceptor, the pH optimum for the serum alpha-L-(1----3)-fucosyltransferase was 6.5. The enzyme was activated by Mn2+ or Mg2+ ions and was inhibited by EDTA. The apparent Km for this enzyme using N-acetyl-2'-O-methyllactosamine was 20.4 mM and Vmax was 5.6 pmol/h/ml serum.
D000606:Amino Sugars / Q000138:chemical synthesis*; D005647:Fucosyltransferases / Q000097:blood*; D006602:Hexosyltransferases / Q000097:blood*; D006801:Humans; D006868:Hydrolysis; D007700:Kinetics; D008970:Molecular Weight; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
D000606:Amino Sugars; C048138:N-acetyl-2'-O-methyllactosamine; D005647:Fucosyltransferases; D006602:Hexosyltransferases; C031692:galactoside 3-fucosyltransferase