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

J. Biol. Chem., Vol. 283, Issue 22, 14971-14979, May 30, 2008

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: 283/22/14971    most recent M709236200v1 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 Google Scholar Google Scholar Articles by Kaneiwa, T. Articles by Sugahara, K. Search for Related Content PubMed PubMed Citation Articles by Kaneiwa, T. Articles by Sugahara, K. Social Bookmarking                      What's this?

Identification of a Novel Chondroitin Hydrolase in Caenorhabditis elegans^*

Tomoyuki Kaneiwa¹, Shuhei Yamada¹², Shuji Mizumoto, Adriana M. Montaño³, Shohei Mitani^¶, and Kazuyuki Sugahara⁴

From the Laboratory of Proteoglycan Signaling and Therapeutics, Hokkaido University Graduate School of Life Science, Sapporo 001-0021, the Department of Biochemistry, Kobe Pharmaceutical University, Higashinada-ku, Kobe 658-8558, and the ^¶Department of Physiology, Tokyo Women's Medical University School of Medicine, Tokyo 162-8666, Japan

Hyaluronidases have been postulated to be the enzyme acting at the initial step of chondroitin sulfate (CS) catabolism in vivo. Since chondroitin (Chn) but not hyaluronic acid (HA) has been detected in Caenorhabditis elegans, the nematode is a good model for elucidating the mechanism of the degradation of CS/Chn in vivo. Here we cloned the homolog of human hyaluronidase in C. elegans, T22C8.2. The Chn-degrading activity in vitro was first demonstrated when it was expressed in COS-7 cells. The enzyme cleaved preferentially Chn. CS-A and CS-C were also depolymerized but to lesser extents, and HA was hardly degraded. In order of preference, the substrates ranked Chn >> CS-A > CS-C >> HA. The products of the degradation of Chn by the enzyme were characterized by anion-exchange high performance liquid chromatography and delayed extraction matrix-assisted laser desorption ionization time-of-flight mass spectrometry. The structure of the major component in the digest was determined as GlcUAβ1-3GalNAcβ1-4GlcUAβ1-3GalNAc, where GlcUA and GalNAc represent D-glucuronic acid and N-acetyl-D-galactosamine, respectively, indicating that this enzyme is a Chn hydrolase, an endo-β-galactosaminidase specific for Chn. Investigation of the effects of pH on the activity revealed the optimum pH of Chn hydrolase to be 6.0. Since Chn in C. elegans has been demonstrated to play critical roles in cell division, Chn hydrolase possibly regulates the function of Chn in vivo. This is the first demonstration of a Chn hydrolase in an animal.

Received for publication, November 9, 2007 , and in revised form, March 25, 2008.

^* The work was supported by HAITEKU (Grant 2004-2008) from the Japan Private School Promotion Foundation, Grant-in-Aid for Scientific Research C-19590052 (to S. Y.) and Scientific Research (B) 18390030 (to K. S.) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, The Human Frontier Science Program Grant GP0018/2005 (to K. S.), and a grant from the Core Research for Evolutional Science and Technology of the Japan Science and Technology agency (to K. S.). 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.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AB361565.

The on-line version of this article (available at http://www.jbc.org) contains two supplemental figures.

¹ Both authors contributed equally to this work.

³ Present address: Doisy Research Center Room 311, Department of Pediatrics, Saint Louis University, 1100 South Grand Blvd., St. Louis, MO 63104.

² To whom correspondence may be addressed: Laboratory of Proteoglycan Signaling and Therapeutics, Graduate School of Life Science, Hokkaido University, Frontier Research Center for Post-Genomic Science and Technology, Nishi 11-choume, Kita 21-jo, Kita-ku, Sapporo, Hokkaido 001-0021, Japan. Tel.: 81-11-706-9055; Fax: 81-11-706-9055; E-mail: tjohej{at}sci.hokudai.ac.jp. ⁴ To whom correspondence may be addressed: Laboratory of Proteoglycan Signaling and Therapeutics, Graduate School of Life Science, Hokkaido University, Frontier Research Center for Post-Genomic Science and Technology, Nishi 11-choume, Kita 21-jo, Kita-ku, Sapporo, Hokkaido 001-0021, Japan. Tel.: 81-11-706-9055; Fax: 81-11-706-9055; E-mail: k-sugar{at}sci.hokudai.ac.jp.

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