title: Characterization of a novel mucin sulphotransferase activity synthesizing sulphated O-glycan core 1,3-sulphate-Gal beta 1-3GalNAc alpha-R.
abstract: A novel sulphotransferase (sulpho-T) activity from rat colonic mucosa was characterized using O-glycan core 1 substrate, Gal beta 1-3GalNAc alpha-benzyl. Derivatives of Gal beta 1-3GalNAc- were used to demonstrate that the 3- and 4-hydroxyl of Gal and the 2-acetamido group of the GalNAc residue of Gal beta 1-3GalNAc alpha-benzyl substrates were important for activity. Sulphated product using Gal beta 1-3GalNAc alpha-benzyl as substrate was analysed by ion spray mass spectrometry, methylation analysis, high-pH anion-exchange chromatography and beta-galactosidase digestion. The results suggested that sulphate was added to the 3-position of the Gal residue. The synthesis of core 2 from core 1 by UDP-GlcNAc: Gal beta 1-3GalNAc beta 6-GlcNAc-transferase was inhibited by sulphation of the Gal residue, indicating that GlcNAc beta 1-6 branching has to precede sulphation in the O-glycan core 1 processing pathway. These data demonstrate several novel pathways in the synthesis of sulphated mucin-type oligosaccharides.
authors.affiliation: Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada.
authors.forename: W
authors.initials: W
authors.lastname: Kuhns
authors.forename: R K
authors.initials: RK
authors.lastname: Jain
authors.forename: K L
authors.initials: KL
authors.lastname: Matta
authors.forename: H
authors.initials: H
authors.lastname: Paulsen
authors.forename: M A
authors.initials: MA
authors.lastname: Baker
authors.forename: R
authors.initials: R
authors.lastname: Geyer
authors.forename: I
authors.initials: I
authors.lastname: Brockhausen
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D002645:Chickens; D003106:Colon / Q000201:enzymology; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000201:enzymology; D007700:Kinetics; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009844:Oligosaccharides / Q000096:biosynthesis* / Q000737:chemistry; D010057:Oviducts / Q000201:enzymology; D011134:Polysaccharides / Q000096:biosynthesis* / Q000737:chemistry; D051381:Rats; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000302:isolation & purification / Q000378:metabolism*; D014018:Tissue Distribution
chemical_list: D009844:Oligosaccharides; D011134:Polysaccharides; D015238:Sulfotransferases; C072220:mucus glycoprotein sulfotransferase