caa a22 4500 469115394 CHVBK 20180323133123.0 cr unu---uuuuu 170328e19921001xx s 000 0 eng 10.1007/BF00731136 doi (NATIONALLICENCE)springer-10.1007/BF00731136 Purification of the Lewis blood-group gene associated α-3/4-fucosyltransferase from human milk: an enzyme transferring fucose primarily to Type 1 and lactose-based oligosaccharide chains [Elektronische Daten] [Philip Johnson, Winifred Watkins] A soluble Lewis blood-group gene associated α-3/4-L-fucosyltransferase has been purified from human milk by a series of steps involving hydrophobic chromatography on Phenyl Sepharose 4B, ion exchange chromatography on CM-Sephadex C-50, affinity chromatography on GDP-hexanolamine Sepharose 4B and gel filtration on Sephacryl S-200. The first step separated α-3-L-fucosyltransferase activity directed towardsN-acetylglucosamine in Type 2 (Galβ1-4GlcNAc-R) acceptors from an α-3/4-fucosyltransferase fraction acting on both Type 1 (Galβ1-3GlcNAc-R) and Type 2 acceptors. Further purification of this latter fraction on CM-Sephadex and GDP-hexanolamine Sepharose gave a single peak of fucosyltransferase activity that catalysed the addition of fucose toN-acetylglucosamine in both Type 1 and Type 2 acceptors and to theO-3 position of glucose in lactose-based oligosaccharides. The enzyme preparation at this stage resembled previously described α-3/4-fucosyltransferase preparations purified from human milk. However, gel filtration of this preparation on Sephacryl S-200 or Sephadex G-150 separated further amounts of α-3-fucosyltransferase activity acting solely on Type 2 acceptors and left a residual α-3/4-fucosyltransferase that retained strong α-4 activity with the Type 1 acceptor, lacto-N-biose 1, and α-3 activity with 2′-fucosyllactose, but had relatively little α-3 activity withN-acetyllactosamine and virtually no capacity to transfer fucose to glycoproteins withN-linked oligosaccharide chains having unsubstituted terminal Type 2 structures. Chapman & Hall, 1992 α-3/4-L-Fucosyltransferase nationallicence α-3- L -fucosyltransferase nationallicence Lewis-gene encoded enzyme nationallicence Lewis antigens nationallicence X-antigen nationallicence human milk nationallicence Johnson Philip Division of Immunochemical Genetics, MRC Clinical Research Centre, Watford Road, HA1 3UJ, Harrow, Middlesex, UK aut Watkins Winifred Division of Immunochemical Genetics, MRC Clinical Research Centre, Watford Road, HA1 3UJ, Harrow, Middlesex, UK aut Glycoconjugate Journal Kluwer Academic Publishers 9/5(1992-10-01), 241-249 0282-0080 9:5<241 1992 9 10719 https://doi.org/10.1007/BF00731136 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00731136 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Johnson Philip Division of Immunochemical Genetics, MRC Clinical Research Centre, Watford Road, HA1 3UJ, Harrow, Middlesex, UK aut NATIONALLICENCE 700 1- Watkins Winifred Division of Immunochemical Genetics, MRC Clinical Research Centre, Watford Road, HA1 3UJ, Harrow, Middlesex, UK aut NATIONALLICENCE 773 0- Glycoconjugate Journal Kluwer Academic Publishers 9/5(1992-10-01), 241-249 0282-0080 9:5<241 1992 9 10719 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer