HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Zhang, L. Articles by Rosenberg, R. D. Search for Related Content PubMed PubMed Citation Articles by Zhang, L. Articles by Rosenberg, R. D.

J Biol Chem, Vol. 274, Issue 9, 5681-5691, February 26, 1999

Anticoagulant Heparan Sulfate Precursor Structures in F9 Embryonal Carcinoma Cells

Lijuan Zhang, Keiichi Yoshida^¶, Jian Liu, and Robert D. Rosenberg

From the  Department of Biology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, the  Department of Medicine, Harvard Medical School, Beth Israel Hospital, Boston, Massachusetts 02215, and ^¶ Tokyo Research Institute of Seikagaku Corp., Higashiyamato-shi, Tokyo 207, Japan

To understand the mechanisms that control anticoagulant heparan sulfate (HS^act) biosynthesis, we previously showed that HS^act production in the F9 system is determined by the abundance of 3-O-sulfotransferase-1 as well as the size of the HS^act precursor pool. In this study, HS^act precursor structures have been studied by characterizing [6-³H]GlcN metabolically labeled F9 HS tagged with 3-O-sulfates in vitro by 3'-phosphoadenosine 5'-phospho-³⁵S and purified 3-O-sulfotransferase-1. This later in vitro labeling allows the regions of HS destined to become the antithrombin (AT)-binding sites to be tagged for subsequent structural studies. It was shown that six 3-O-sulfation sites exist per HS^act precursor chain. At least five out of six 3-O-sulfate-tagged oligosaccharides in HS^act precursors bind AT, whereas none of 3-O-sulfate-tagged oligosaccharides from HS^inact precursors bind AT. When treated with low pH nitrous or heparitinase, 3-O-sulfate-tagged HS^act and HS^inact precursors exhibit clearly different structural features. 3-O-Sulfate-tagged HS^act hexasaccharides were AT affinity purified and sequenced by chemical and enzymatic degradations. The 3-O-sulfate-tagged HS^act hexasaccharides exhibited the following structures, UA-[6-³H]GlcNAc6S-GlcUA-[6-³H]GlcNS3³⁵S±6S-IdceA2S-[6-³H]GlcNS6S. The underlined 6- and 3-O-sulfates constitute the most critical groups for AT binding in view of the fact that the precursor hexasaccharides possess all the elements for AT binding except for the 3-O-sulfate moiety. The presence of five potential AT-binding precursor hexasaccharides in all HS^act precursor chains demonstrates for the first time the processive assembly of specific sequence in HS. The difference in structures around potential 3-O-sulfate acceptor sites in HS^act and HS^inact precursors suggests that these precursors might be generated by different concerted assembly mechanisms in the same cell. This study permits us to understand better the nature of the HS biosynthetic pathway that leads to the generation of specific saccharide sequences.

---

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.

This article has been cited by other articles:

				E. P. Girardin, S. HajMohammadi, B. Birmele, A. Helisch, N. W. Shworak, and A. I. de Agostini Synthesis of Anticoagulantly Active Heparan Sulfate Proteoglycans by Glomerular Epithelial Cells Involves Multiple 3-O-Sulfotransferase Isoforms and a Limiting Precursor Pool J. Biol. Chem., November 11, 2005; 280(45): 38059 - 38070. [Abstract] [Full Text] [PDF]

				M Contini, S Pacini, L Ibba-Manneschi, V Boddi, M Ruggiero, G Liguri, M Gulisano, C Catini, and P A S Mourao Modification of plasma glycosaminoglycans in long distance runners * Commentary Br. J. Sports Med., April 1, 2004; 38(2): 134 - 137. [Abstract] [Full Text] [PDF]

				Z. L. Wu, M. Lech, D. L. Beeler, and R. D. Rosenberg Determining Heparan Sulfate Structure in the Vicinity of Specific Sulfotransferase Recognition Sites by Mass Spectrometry J. Biol. Chem., January 16, 2004; 279(3): 1861 - 1866. [Abstract] [Full Text] [PDF]

				J. Chen, M. B. Duncan, K. Carrick, R. M. Pope, and J. Liu Biosynthesis of 3-O-sulfated heparan sulfate: unique substrate specificity of heparan sulfate 3-O-sulfotransferase isoform 5 Glycobiology, November 1, 2003; 13(11): 785 - 794. [Abstract] [Full Text] [PDF]

				G. Xia, J. Chen, V. Tiwari, W. Ju, J.-P. Li, A. Malmstrom, D. Shukla, and J. Liu Heparan Sulfate 3-O-Sulfotransferase Isoform 5 Generates Both an Antithrombin-binding Site and an Entry Receptor for Herpes Simplex Virus, Type 1 J. Biol. Chem., September 27, 2002; 277(40): 37912 - 37919. [Abstract] [Full Text] [PDF]

				J. Liu, Z. Shriver, R. M. Pope, S. C. Thorp, M. B. Duncan, R. J. Copeland, C. S. Raska, K. Yoshida, R. J. Eisenberg, G. Cohen, et al. Characterization of a Heparan Sulfate Octasaccharide That Binds to Herpes Simplex Virus Type 1 Glycoprotein D J. Biol. Chem., August 30, 2002; 277(36): 33456 - 33467. [Abstract] [Full Text] [PDF]

				S. Hasan, G. Hosseini, M. Princivalle, J.-C. Dong, D. Birsan, C. Cagide, and A. I. de Agostini Coordinate Expression of Anticoagulant Heparan Sulfate Proteoglycans and Serine Protease Inhibitors in the Rat Ovary: A Potent System of Proteolysis Control Biol Reprod, January 1, 2002; 66(1): 144 - 158. [Abstract] [Full Text]

				L. Zhang, D. L. Beeler, R. Lawrence, M. Lech, J. Liu, J. C. Davis, Z. Shriver, R. Sasisekharan, and R. D. Rosenberg 6-O-Sulfotransferase-1 Represents a Critical Enzyme in the Anticoagulant Heparan Sulfate Biosynthetic Pathway J. Biol. Chem., November 2, 2001; 276(45): 42311 - 42321. [Abstract] [Full Text] [PDF]

				M. Princivalle, S. Hasan, G. Hosseini, and A. I. de Agostini Anticoagulant heparan sulfate proteoglycans expression in the rat ovary peaks in preovulatory granulosa cells Glycobiology, March 1, 2001; 11(3): 183 - 194. [Abstract] [Full Text] [PDF]

				J. Liu, Z. Shriver, P. Blaiklock, K. Yoshida, R. Sasisekharan, and R. D. Rosenberg Heparan Sulfate D-Glucosaminyl 3-O-Sulfotransferase-3A Sulfates N-Unsubstituted Glucosamine Residues J. Biol. Chem., December 31, 1999; 274(53): 38155 - 38162. [Abstract] [Full Text] [PDF]

				W. L. McKeehan, X. Wu, and M. Kan Requirement for Anticoagulant Heparan Sulfate in the Fibroblast Growth Factor Receptor Complex J. Biol. Chem., July 30, 1999; 274(31): 21511 - 21514. [Abstract] [Full Text] [PDF]

				L. Zhang, R. Lawrence, J. J. Schwartz, X. Bai, G. Wei, J. D. Esko, and R. D. Rosenberg The Effect of Precursor Structures on the Action of Glucosaminyl 3-O-Sulfotransferase-1 and the Biosynthesis of Anticoagulant Heparan Sulfate J. Biol. Chem., July 27, 2001; 276(31): 28806 - 28813. [Abstract] [Full Text] [PDF]