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

J. Biol. Chem., Vol. 281, Issue 19, 13644-13651, May 12, 2006

This Article Full Text Full Text (PDF) All Versions of this Article: 281/19/13644    most recent M601453200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted 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 Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Kizuka, Y. Articles by Oka, S. Search for Related Content PubMed PubMed Citation Articles by Kizuka, Y. Articles by Oka, S. Social Bookmarking                      What's this?

Physical and Functional Association of Glucuronyltransferases and Sulfotransferase Involved in HNK-1 Biosynthesis^*

Yasuhiko Kizuka, Takahiro Matsui, Hiromu Takematsu, Yasunori Kozutsumi, Toshisuke Kawasaki^¶, and Shogo Oka¹

From the Department of Biological Chemistry, Graduate School of Pharmaceutical Sciences and Laboratory of Membrane Biochemistry and Biophysics, Graduate School of Biostudies, Kyoto University, Kyoto 606-8501 and ^¶Research Center for Glycobiotechnology, Ritsumeikan University, Shiga 525-8577, Japan

HNK-1 carbohydrate expressed predominantly in the nervous system is considered to be involved in cell migration, recognition, adhesion, and synaptic plasticity. Human natural killer-1 (HNK-1) carbohydrate has a unique structure consisting of a sulfated trisaccharide (HSO₃-3GlcA1-3Gal1-4GlcNAc-) and is sequentially biosynthesized by one of two glucuronyltransferases (GlcAT-P or GlcAT-S) and a sulfotransferase (HNK-1ST). Considering that almost all the HNK-1 carbohydrate structures so far determined in the nervous system are sulfated, we hypothesized that GlcAT-P or GlcAT-S functionally associates with HNK-1ST, which results in efficient sequential biosynthesis of HNK-1 carbohydrate. In this study, we demonstrated that both GlcAT-P and GlcAT-S were co-immunoprecipitated with HNK-1ST with a transient expression system in Chinese hamster ovary cells. Immunofluorescence staining revealed that these enzymes are mainly co-localized in the Golgi apparatus. To determine which domain is involved in this interaction, we prepared the C-terminal catalytic domains of GlcAT-P, GlcAT-S, and HNK-1ST, and we then performed pulldown assays with the purified enzymes. As a result, we obtained evidence that mutual catalytic domains of GlcAT-P or GlcAT-S and HNK-1ST are important and sufficient for formation of an enzyme complex. With an in vitro assay system, the activity of HNK-1ST increased about 2-fold in the presence of GlcAT-P or GlcAT-S compared with that in its absence. These results suggest that the function of this enzyme complex is relevant to the efficient sequential biosynthesis of the HNK-1 carbohydrate.

Received for publication, February 14, 2006 , and in revised form, March 16, 2006.

^* This work was supported in part by a Grant-in-aid for Creative Scientific Research 16GS0313 (to S. O.) and a Grant-in-aid for Scientific Research on Priority Areas A-14082203 (to T. K.) from the Ministry of Education, Culture, Sports, and Technology. 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: Dept. of Biological Chemistry, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan. Tel.: 81-75-753-4562; Fax: 81-75-753-4605; E-mail: shogo{at}pharm.kyoto-u.ac.jp.

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

This article has been cited by other articles:

				X. Zhao, C. Graves, S. J. Ames, D. E. Fisher, and R. A. Spanjaard Mechanism of Regulation and Suppression of Melanoma Invasiveness by Novel Retinoic Acid Receptor-{gamma} Target Gene Carbohydrate Sulfotransferase 10 Cancer Res., June 15, 2009; 69(12): 5218 - 5225. [Abstract] [Full Text] [PDF]

				Y. Kizuka, Y. Tonoyama, and S. Oka Distinct Transport and Intracellular Activities of Two GlcAT-P Isoforms J. Biol. Chem., April 3, 2009; 284(14): 9247 - 9256. [Abstract] [Full Text] [PDF]

				H. Miller-Podraza, K. Weikkolainen, T. Larsson, P. Linde, J. Helin, J. Natunen, and K.-A. Karlsson Helicobacter pylori binding to new glycans based on N-acetyllactosamine Glycobiology, April 1, 2009; 19(4): 399 - 407. [Abstract] [Full Text] [PDF]

				I. Morita, Y. Kizuka, S. Kakuda, and S. Oka Expression and Function of the HNK-1 Carbohydrate J. Biochem., June 1, 2008; 143(6): 719 - 724. [Abstract] [Full Text] [PDF]