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

This Article 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 Kusche, M. Articles by Lindahl, U. Search for Related Content PubMed PubMed Citation Articles by Kusche, M. Articles by Lindahl, U. Social Bookmarking                      What's this?

J. Biol. Chem., Vol. 263, Issue 30, 15474-15484, 10, 1988

Biosynthesis of heparin. O-sulfation of the antithrombin-binding region

M Kusche, G Backstrom, J Riesenfeld, M Petitou, J Choay and U Lindahl
Department of Veterinary Medical Chemistry, Swedish University of Agricultural Sciences, Uppsala, Sweden.

The antithrombin-binding region in heparin is a pentasaccharide sequence with the predominant structure GlcNAc(6-OSO3)-GlcA-GlcNSO3(3,6- di-OSO3)-IdoA -(2-OSO3)-GlcNSO3(6-OSO3) (where GlcA and IdoA represent D-glucuronic and L-iduronic acid, respectively), in which the 3-O- sulfate residue on the internal glucosaminyl unit is a marker group for this particular region of the polysaccharide molecule. A heparin octasaccharide which contained the above pentasaccharide sequence was N/O-desulfated and re-N-sulfated and was then incubated with adenosine 3'-phosphate 5'-phospho[35S]sulfate in the presence of a microsomal fraction from mouse mastocytoma tissue. Fractionation of the resulting 35S-labeled octasaccharide on antithrombin-Sepharose yielded a high affinity fraction that accounted for approximately 2% of the total incorporated label. Structural analysis of this fraction indicated that the internal glucosamine unit of the pentasaccharide sequence was 3-O- 35S-sulfated, whereas both adjacent glucosamine units carried 6-O- [35S]sulfate groups. In contrast, the fractions with low affinity for antithrombin (approximately 98% of incorporated 35S) showed no consistent O-35S sulfation pattern and essentially lacked glucosaminyl 3-O-[35S]sulfate groups. It is suggested that the 3-O-sulfation reaction concludes the formation of the antithrombin-binding region. This proposal was corroborated in a similar experiment using a synthetic pentasaccharide with the structure GlcNSO3(6-OSO3)-GlcA- GlcNSO3(6-OSO3)-Id oA (2-OSO3)-GlcNSO3(6-OSO3) as sulfate acceptor. This molecule corresponds to a functional antithrombin-binding region but for the lack of a 3-O-sulfate group at the internal glucosamine unit. The 35S-labeled pentasaccharide recovered after incubation bound with high affinity to antithrombin-Sepharose and contained a 3-O- [35S]sulfate group at the internal glucosamine residue as the only detectable labeled component. The use of this pentasaccharide substrate along with the affinity matrix provides a highly specific assay for the 3-O-sulfotransferase.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				R. Lawrence, B. Kuberan, M. Lech, D. L. Beeler, and R. D. Rosenberg Mapping critical biological motifs and biosynthetic pathways of heparan sulfate Glycobiology, May 1, 2004; 14(5): 467 - 479. [Abstract] [Full Text] [PDF]

				D. S. Pikas, J.-p. Li, I. Vlodavsky, and U. Lindahl Substrate Specificity of Heparanases from Human Hepatoma and Platelets J. Biol. Chem., July 24, 1998; 273(30): 18770 - 18777. [Abstract] [Full Text] [PDF]

				G. C. Jayson, M. Lyon, C. Paraskeva, J. E. Turnbull, J. A. Deakin, and J. T. Gallagher Heparan Sulfate Undergoes Specific Structural Changes during the Progression from Human Colon Adenoma to Carcinoma in Vitro J. Biol. Chem., January 2, 1998; 273(1): 51 - 57. [Abstract] [Full Text] [PDF]

				L. Uhlin-Hansen, M. Kusche-Gullberg, E. Berg, I. Eriksson, and L. Kjellen Mouse Mastocytoma Cells Synthesize Undersulfated Heparin and Chondroitin Sulfate in the Presence of Brefeldin A J. Biol. Chem., February 7, 1997; 272(6): 3200 - 3206. [Abstract] [Full Text] [PDF]

				J. Liu, N. W. Shworak, L. M.S. Fritze, J. M. Edelberg, and R. D. Rosenberg Purification of Heparan Sulfate D-Glucosaminyl 3-O-Sulfotransferase J. Biol. Chem., October 25, 1996; 271(43): 27072 - 27082. [Abstract] [Full Text] [PDF]

				H. Tsuda, S. Yamada, Y. Yamane, K. Yoshida, J. J. Hopwood, and K. Sugahara Structures of Five Sulfated Hexasaccharides Prepared from Porcine Intestinal Heparin Using Bacterial Heparinase J. Biol. Chem., May 3, 1996; 271(18): 10495 - 10502. [Abstract] [Full Text] [PDF]

				M. Kobayashi, H. Habuchi, O. Habuchi, M. Saito, and K. Kimata Purification and Characterization of Heparan Sulfate 2-Sulfotransferase from Cultured Chinese Hamster Ovary Cells J. Biol. Chem., March 29, 1996; 271(13): 7645 - 7653. [Abstract] [Full Text] [PDF]

				H. Habuchi, O. Habuchi, and K. Kimata Purification and Characterization of Heparan Sulfate 6-Sulfotransferase from the Culture Medium of Chinese Hamster Ovary Cells J. Biol. Chem., February 24, 1995; 270(8): 4172 - 4179. [Abstract] [Full Text] [PDF]