title: Determination of substrate specificity of sulfotransferases and glycosyltransferases (proteoglycans).
abstract: Proteoglycans have sulfated linear polysaccharide chains, that is, heparan sulfate, heparin, chondroitin sulfates, dermatan sulfate, and keratan sulfate. Many glycosyltransferases and sulfotransferases are involved in biosynthesis of the polysaccharides. Specificities of these enzymes have been mainly determined by evaluating their activities to various acceptor carbohydrates and by analyzing the structure of the products. For the latter purpose, enzymatic hydrolysis using heparitinases, heparinase, and chondroitinases or chemical degradation employing nitrous acid deamination has been effectively used in combination with high-performance liquid chromatography (HPLC) of the degraded products. As examples, we describe methods for assays and product characterization of sulfotransferases involved in biosynthesis of these polysaccharides, namely heparan sulfate 2-sulfotransferase, heparan sulfate 6-sulfotransferases, chondroitin 4-sulfotransferases, chondroitin 6-sulfotransferase, N-acetylgalactosamine 4-sulfate 6-sulfotransferase, and N-acetylglucosamine 6-sulfotransferases.
authors.affiliation: Institute for Molecular Science of Medicine, Aichi Medical University, Nagakute, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Habuchi
authors.forename: Osami
authors.initials: O
authors.lastname: Habuchi
authors.forename: Kenji
authors.initials: K
authors.lastname: Uchimura
authors.forename: Koji
authors.initials: K
authors.lastname: Kimata
authors.forename: Takashi
authors.initials: T
authors.lastname: Muramatsu
mesh_terms: D003641:Deamination; D016695:Glycosyltransferases / Q000502:physiology*; D006868:Hydrolysis; D009608:Nitrous Acid / Q000737:chemistry; D011509:Proteoglycans / Q000096:biosynthesis*; D013379:Substrate Specificity; D015238:Sulfotransferases / Q000502:physiology*
chemical_list: D011509:Proteoglycans; D016695:Glycosyltransferases; D015238:Sulfotransferases; D009608:Nitrous Acid