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Reassessment of the acceptor specificity and general properties of the Lewis blood-group gene associated α-3/4-fucosyltransferase purified from human milk

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Abstract

The acceptor specificity and general properties of a Lewis blood-group gene associated α-3/4-L-fucosyltransferase isolated from human milk have been examined at the penultimate purification stage involving affinity chromatography on GDP-hexanolamine Sepharose, and after a subsequent gel filtration step on Sephacryl S-200. Both preparations transferred fucose to theO-4 position ofN-acetylglucosamine in Type 1 (Galβ1-3GlcNAc-R) acceptors and theO-3 position of glucose in lactose-based (Galβ1-4Glc) oligosaccharides, and both used Type 1 sialylated compounds when the terminalN-acetylneuraminic acid was present in α-2,3 linkage. The striking difference between the two preparations was in their reactivity with Type 2 (Galβ1-4GlcNAc-R) chains; after Sephacryl S-200 chromatography the apparentK M values for the α-3/4- preparation with unsubstituted low-molecular-weight Type 2 oligosaccharides were considerably increased. Substitution of the terminal galactose with sialic acid in α-2,3 linkage decreased theK M values for low-molecular-weight oligosaccharides but no detectable incorporation of fucose was observed intoN-acetyllactosamine end-groups of glycoproteins withN-linked oligosaccharide chains, irrespective of the presence of sialic acid in the terminal sequences.

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Authors and Affiliations

  1. Division of Immunochemical Genetics, MRC Clinical Research Centre, Watford Road, HA1 3UJ, Harrow, Middlesex, UK

    Philip H. Johnson, Alastair S. R. Donald & Winifred M. Watkins

  2. MRC Human Biochemical Genetics Unit, University College London, Wolfson House, 4 Stephenson Way, NW1 2HE, London, UK

    Philip H. Johnson

  3. Biomedical Nuclear Magnetic Resonance Centre, National Institute for Medical Research, Mill Hill, NW7 1AA, London, UK

    James Feeney

  4. Department of Haematology, Royal Postgraduate Medical School, Hammersmith Hospital, Du Cane Road, W12 0NN, London, UK

    Winifred M. Watkins

Authors
  1. Philip H. Johnson
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  2. Alastair S. R. Donald
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  3. James Feeney
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  4. Winifred M. Watkins
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Deceased 25 June 1991.

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Johnson, P.H., Donald, A.S.R., Feeney, J. et al. Reassessment of the acceptor specificity and general properties of the Lewis blood-group gene associated α-3/4-fucosyltransferase purified from human milk. Glycoconjugate J 9, 251–264 (1992). https://doi.org/10.1007/BF00731137

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  • DOI: https://doi.org/10.1007/BF00731137

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