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Synthesis of the acceptor analog αFuc(1→2)αGal-O(CH2)7 CH3: A probe for the kinetic mechanism of recombinant human blood group B glycosyltransferase

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Abstract

We report the chemical synthesis of αFuc(1→2)αGal-O(CH2)7CH3 (1) an analog of the natural blood group (O)H disaccharide αFuc(1→2)βGal-OR. Compound 1 was a good substrate for recombinant blood group B glycosyltransferase (GTB) and was used as a precursor for the enzymatic synthesis of the blood group B analog αGal(→3)[αFuc(1→2)]αGal-O(CH2)7CH3 (2). To probe the mechanism of the GTB reaction, kinetic evaluations were carried out employing compound 1 or the natural acceptor disaccharide αFuc(1→2)βGal-O(CH2)7CH3 (3) with UDP-Gal and UDP-GalNAc donors. Comparisons of the kinetic constants for alternative donor and acceptor pairs suggest that the GTB mechanism is Theorell-Chance where donor binding precedes acceptor binding. GTB operates with retention of configuration at the anomeric center of the donor. Retaining reactions are thought to occur via a double-displacement mechanism with formation of a glycosyl-enzyme intermediate consistent with the proposed Theorell-Chance mechanism.

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Authors and Affiliations

  1. Department of Chemistry, University of Alberta, Edmonton, Alberta, T6G 2G2, Canada

    Vivekanand P. Kamath, Catharine A. Compston, Ole Hindsgaul & Monica M. Palcic

  2. Institute for Biological Sciences, National Research Council of Canada, Ottawa, Ontario, K1A 0R6, Canada

    Nina O.L. Seto

Authors
  1. Vivekanand P. Kamath
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  2. Nina O.L. Seto
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  3. Catharine A. Compston
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  4. Ole Hindsgaul
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  5. Monica M. Palcic
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Kamath, V.P., Seto, N.O., Compston, C.A. et al. Synthesis of the acceptor analog αFuc(1→2)αGal-O(CH2)7 CH3: A probe for the kinetic mechanism of recombinant human blood group B glycosyltransferase. Glycoconj J 16, 599–606 (1999). https://doi.org/10.1023/A:1007072832421

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