title: Purification and characterization of N-acetylglucosaminyl sulfotransferase from chick corneas.
abstract: N-Acetylglucosaminyl(GlcNAc) sulfotransferase, which transfers sulfate from adenosine 3'-phosphate 5'-phosphosulfate to GlcNAc at the nonreducing end of oligosaccharides, was purified 887-fold with a 8.4% yield from 2-day-old chick corneas by chromatography on CM-Sepharose, WGA-agarose, GlcNAc-agarose, and 3',5'-ADP-agarose columns. The purified enzyme has an optimum pH of 6.0 (Mes buffer) and specifically transfers a sulfate to GlcNAc at the nonreducing end but not to internal GlcNAc. The enzyme was stimulated by protamine and Mn(2+). SDS-polyacrylamide gel electrophoresis of the purified enzyme still showed two main bands (66 and 55 kDa) with some minor bands. It appears that this enzyme competes with beta-galactosyltransferase in binding to the nonreducing GlcNAc residue on KS synthesis; this suggests that the sulfation of the GlcNAc residue is coupled with the elongation of the KS chain.
authors.affiliation: Section of Radiochemistry, Faculty of Pharmacy, Meijo University, 150 Yagotoyama, Tempaku-ku, Nagoya 468-8503, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: I
authors.initials: I
authors.lastname: Takahashi
authors.forename: N
authors.initials: N
authors.lastname: Ogata
authors.forename: K
authors.initials: K
authors.lastname: Nakazawa
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D002240:Carbohydrate Sequence; D002645:Chickens; D002809:Chondroitin Sulfates / Q000737:chemistry / Q000378:metabolism; D002847:Chromatography, Agarose; D003315:Cornea / Q000201:enzymology*; D003871:Dermatan Sulfate / Q000737:chemistry / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007632:Keratan Sulfate / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D000097811:Carbohydrate Sulfotransferases
chemical_list: D002809:Chondroitin Sulfates; D003871:Dermatan Sulfate; D007632:Keratan Sulfate; D015238:Sulfotransferases; D000097811:Carbohydrate Sulfotransferases