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

J. Biol. Chem., Vol. 282, Issue 51, 37134-37145, December 21, 2007

This Article Full Text Full Text (PDF) All Versions of this Article: 282/51/37134    most recent M704663200v1 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 Ochiai, A. Articles by Murata, K. Search for Related Content PubMed PubMed Citation Articles by Ochiai, A. Articles by Murata, K. Social Bookmarking                      What's this?

A Novel Structural Fold in Polysaccharide Lyases

BACILLUS SUBTILIS FAMILY 11 RHAMNOGALACTURONAN LYASE YesW WITH AN EIGHT-BLADED β-PROPELLER^*

Akihito Ochiai, Takafumi Itoh, Yukie Maruyama, Akiko Kawamata, Bunzo Mikami, Wataru Hashimoto, and Kousaku Murata¹

From the Laboratory of Basic and Applied Molecular Biotechnology and the Laboratory of Applied Structural Biology, Graduate School of Agriculture, Kyoto University, Uji, Kyoto 611-0011, Japan

Rhamnogalacturonan (RG) lyase produced by plant pathogenic and saprophytic microbes plays an important role in degrading plant cell walls. An extracellular RG lyase YesW from saprophytic Bacillus subtilis is a member of polysaccharide lyase family 11 and cleaves glycoside bonds in polygalacturonan as well as RG type-I through a β-elimination reaction. Crystal structures of YesW and its complex with galacturonan disaccharide, a reaction product analogue, were determined at 1.4 and 2.5Å resolutions with final R-factors of 16.4% and 16.6%, respectively. The enzyme is composed of an eight-bladed β-propeller with a deep cleft in the center as a basic scaffold, and its structural fold has not been seen in polysaccharide lyases analyzed thus far. Structural analysis of the disaccharide-bound YesW and a site-directed mutagenesis study suggested that Arg-452 and Lys-535 stabilize the carboxyl group of the acidic polysaccharide molecule and Tyr-595 makes a stack interaction with the sugar pyranose ring. In addition to amino acid residues binding to the disaccharide, one calcium ion, which is coordinated by Asp-401, Glu-422, His-363, and His-399, may mediate the enzyme activity. This is, to our knowledge, the first report of a new structural category with a β-propeller fold in polysaccharide lyases and provides structural insights into substrate binding by RG lyase.

Received for publication, June 6, 2007 , and in revised form, October 3, 2007.

The atomic coordinates and structure factors (code 2Z8R and 2Z8S) have been deposited in the Protein Data Bank, Research Collaboratory for Structural Bioinformatics, Rutgers University, New Brunswick, NJ (http://www.rcsb.org/).

^* This work was supported in part by Grants-in-aid from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (to K. M. and W. H.), by COE for Microbial-Process Development Pioneering Future Production Systems from the Ministry of Education, Culture, Sports, Science, and Technology of Japan, and by the Targeted Proteins Research Program from the Ministry of Education, Culture, Sports, Science, and Technology of Japan. Part of this work was supported by Research Fellowships from the Japan Society for the Promotion of Science for Young Scientists (to A. O.). 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.: 81-774-38-3766; Fax: 81-774-38-3767; E-mail: kmurata{at}kais.kyoto-u.ac.jp.

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

This article has been cited by other articles:

				A. Ochiai, T. Itoh, B. Mikami, W. Hashimoto, and K. Murata Structural Determinants Responsible for Substrate Recognition and Mode of Action in Family 11 Polysaccharide Lyases J. Biol. Chem., April 10, 2009; 284(15): 10181 - 10189. [Abstract] [Full Text] [PDF]