title: Biosynthesis of heparin. Different molecular forms of O-sulfotransferases.
abstract: O-Sulfotransferases involved in heparin biosynthesis were purified > or = 10,000-fold from detergent extracts of mouse mastocytoma tissue by sequential chromatographies on DEAE-Sephacel, heparin-agarose, blue Sepharose, and 3',5'-ADP-Sepharose. The resultant preparation catalyzed the transfer of 35S from 3'-phosphoadenosyl-5'-phospho-[35S]sulfate into N,O-desulfated, re-N-sulfated heparin. Anion-exchange high performance liquid chromatography of disaccharides obtained by deaminative cleavage of the 35S-labeled polysaccharide product revealed O-35S-sulfation at C-2 of L-iduronic acid and at C-6 of D-glucosamine units. SDS-polyacrylamide gel electrophoresis of semipurified enzyme followed by extraction of gel segments and renaturation of proteins consistently showed two distinct fractions of O-sulfotransferase activity, corresponding to proteins of approximately 20 and approximately 60 kDa. The approximately 60-kDa enzyme(s) catalyzed both the 2-O- and 6-O-sulfotransferase reactions, whereas the approximately 20-kDa fraction promoted iduronosyl 2-O-sulfation only. These results are discussed in relation to previous findings, indicating that some of the enzymes involved in heparin biosynthesis catalyze more than one reaction.
authors.affiliation: Department of Medical and Physiological Chemistry, Uppsala University, Sweden.
authors.forename: H
authors.initials: H
authors.lastname: Wlad
authors.forename: M
authors.initials: M
authors.lastname: Maccarana
authors.forename: I
authors.initials: I
authors.lastname: Eriksson
authors.forename: L
authors.initials: L
authors.lastname: Kjellén
authors.forename: U
authors.initials: U
authors.lastname: Lindahl
mesh_terms: D000818:Animals; D002240:Carbohydrate Sequence; D006493:Heparin / Q000096:biosynthesis*; D012515:Mast-Cell Sarcoma / Q000201:enzymology; D051379:Mice; D008969:Molecular Sequence Data; D015238:Sulfotransferases / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D006493:Heparin; D015238:Sulfotransferases