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

J. Biol. Chem., Vol. 280, Issue 47, 39115-39123, November 25, 2005

This Article Full Text Full Text (PDF) All Versions of this Article: 280/47/39115    most recent M508816200v1 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 Ohtake, S. Articles by Habuchi, O. Search for Related Content PubMed PubMed Citation Articles by Ohtake, S. Articles by Habuchi, O. Social Bookmarking                      What's this?

Recognition of Sulfation Pattern of Chondroitin Sulfate by Uronosyl 2-O-Sulfotransferase^*

Shiori Ohtake, Koji Kimata, and Osami Habuchi1

From the Department of Chemistry, Aichi University of Education, Igaya-cho, Kariya, Aichi 448-8542 and the Institute for Molecular Science of Medicine, Aichi Medical University, Nagakute, Aichi 480-1195, Japan

We have shown previously that a highly sulfated sequence, GalNAc(4,6-SO₄)-GlcA(2SO₄)-GalNAc(6SO₄), is present at the nonreducing terminal of chondroitin sulfate (CS), and this structure was synthesized from a unique sequence, GalNAc(4SO₄)-GlcA(2SO₄)-GalNAc(6SO₄), by sulfation with N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase. Uronosyl 2-O-sulfotrasferase (2OST), which transfers sulfate from 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to position 2 of the GlcA residue of CS, is expected to be involved in synthesis of these structures; however, the specificity of 2OST concerning recognition of the sulfation pattern of the acceptor has largely remained unclear. In the present study, we examined the specificity of 2OST in terms of recognition of the sulfation pattern around the targeting GlcA residue. The recombinant 2OST could sulfate CS-A, CS-C, and desulfated dermatan sulfate. When [³⁵S]glycosaminoglycans formed from CS-A after the reaction with the recombinant 2OST and [³⁵S]PAPS were subjected to limited digestion with chondroitinase ACII, a radioactive tetrasaccharide (Tetra A) was obtained as a sole intermediate product. The sequence of Tetra A was found to be HexA-GalNAc(4SO₄)-GlcA(2SO₄)-GalNAc(6SO₄) by enzymatic and chemical reactions. These observations indicate that 2OST transfers sulfate preferentially to the GlcA residue located in a unique sequence, -GalNAc(4SO₄)-GlcA-GalNAc(6SO₄)-. When oligosaccharides with different sulfation patterns were used as the acceptor, GalNAc(4SO₄)-GlcA-GalNAc(6SO₄) and GlcA-GalNAc(4SO₄)-GlcA-GalNAc(6SO₄) were the best acceptors for 2OST among trisaccharides and tetrasaccharides, respectively. These results suggest that 2OST may be involved in the synthesis of the highly sulfated structure found in CS-A.

Received for publication, August 10, 2005 , and in revised form, September 19, 2005.

^* This work was supported by Grants 14082206 and 16-4208 from the Ministry of Education, Culture, Sports, Science and Technology of Japan, and by a special research fund from the Seikagaku Corporation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed. Tel.: 81-566-26-2642; Fax: 81-566-26-2649; E-mail: ohabuchi{at}auecc.aichi-edu.ac.jp.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				T. Yamaguchi, S. Ohtake, K. Kimata, and O. Habuchi Molecular Cloning of Squid N-Acetylgalactosamine 4-Sulfate 6-O-Sulfotransferase and Synthesis of a Unique Chondroitin Sulfate Containing E D Hybrid Tetrasaccharide Structure by the Recombinant Enzyme Glycobiology, December 1, 2007; 17(12): 1365 - 1376. [Abstract] [Full Text] [PDF]

				D. Xu, D. Song, L. C. Pedersen, and J. Liu Mutational Study of Heparan Sulfate 2-O-Sulfotransferase and Chondroitin Sulfate 2-O-Sulfotransferase J. Biol. Chem., March 16, 2007; 282(11): 8356 - 8367. [Abstract] [Full Text] [PDF]

				A. Yusa, K. Kitajima, and O. Habuchi N-Linked Oligosaccharides on Chondroitin 6-Sulfotransferase-1 Are Required for Production of the Active Enzyme, Golgi Localization, and Sulfotransferase Activity toward Keratan Sulfate J. Biol. Chem., July 21, 2006; 281(29): 20393 - 20403. [Abstract] [Full Text] [PDF]