title: The biosynthesis of anticoagulant heparan sulfate by the heparan sulfate 3-O-sulfotransferase isoform 5.
abstract: The role of heparan sulfate (HS) in regulating blood coagulation has a wide range of clinical implications. In this study, we investigated the role of 3-O-sulfotransferase isoform 5 (3-OST-5) in generating anticoagulant HS in vivo. A Chinese hamster ovary cell line (3OST5/CHO) stably expressing 3-OST-5 was generated. The expression of 3-OST-5 in 3OST5/CHO cells was confirmed by Northern blot analysis, RT-PCR, and the disaccharide analyses of the HS from the cells. We also determined the effects of the HS from 3OST5/CHO on antithrombin-mediated inhibition of factor Xa. Fluorescently labeled antithrombin bound to the surface of 3OST5/CHO cells, suggesting that the antithrombin-binding HS is indeed present on the cell surface. Our results demonstrate that the 3-OST-5 gene is capable of synthesizing anticoagulant HS in CHO cells and has the potential to contribute to the biosynthesis of anticoagulant HS in humans.
authors.affiliation: Division of Medicinal Chemistry and Natural Products, Rm. 309 Beard Hall, School of Pharmacy, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Duncan
authors.forename: Jinghua
authors.initials: J
authors.lastname: Chen
authors.forename: Jeffrey P
authors.initials: JP
authors.lastname: Krise
authors.forename: Jian
authors.initials: J
authors.lastname: Liu
mesh_terms: D000818:Animals; D000925:Anticoagulants / Q000737:chemistry / Q000378:metabolism*; D000991:Antithrombins / Q000378:metabolism; D016466:CHO Cells; D006224:Cricetinae; D004187:Disaccharides / Q000032:analysis; D004586:Electrophoresis; D015951:Factor Xa / Q000378:metabolism; D006497:Heparan Sulfate / Q000096:biosynthesis* / Q000737:chemistry; D006801:Humans; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D015238:Sulfotransferases / Q000235:genetics / Q000378:metabolism*
chemical_list: D000925:Anticoagulants; D000991:Antithrombins; D004187:Disaccharides; D015951:Factor Xa; D006497:Heparan Sulfate; D007527:Isoenzymes; D015238:Sulfotransferases; C021350:heparitin sulfotransferase