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J. Biol. Chem., Vol. 283, Issue 26, 18260-18268, June 27, 2008

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The Crystal Structure of Pectate Lyase PelI from Soft Rot Pathogen Erwinia chrysanthemi in Complex with Its Substrate^*

Christophe Creze, Sandra Castang¹, Emmanuel Derivery², Richard Haser, Nicole Hugouvieux-Cotte-Pattat, Vladimir E. Shevchik, and Patrice Gouet³

From the Laboratoire de BioCristallographie, Institut de Biologie et Chimie des Protéines, CNRS et Université de Lyon, UMR 5086, IFR 128 "BioSciences Gerland-Lyon Sud", F-69367 Lyon Cedex 07 and Unité Microbiologie Adaptation et Pathogénie, UMR 5240 CNRS, Université Lyon 1, F-69622, INSA-Lyon, F-69621 Villeurbanne, France

The crystallographic structure of the family 3 polysaccharide lyase (PL-3) PelI from Erwinia chrysanthemi has been solved to 1.45Å resolution. It consists of an N-terminal domain harboring a fibronectin type III fold linked to a catalytic domain displaying a parallel β-helix topology. The N-terminal domain is located away from the active site and is not involved in the catalytic process. After secretion in planta, the two domains are separated by E. chrysanthemi proteases. This event turns on the hypersensitive response of the host. The structure of the single catalytic domain determined to 2.1Å resolution shows that the domain separation unveils a "Velcro"-like motif of asparagines, which might be recognized by a plant receptor. The structure of PelI in complex with its substrate, a tetragalacturonate, has been solved to 2.3Å resolution. The sugar binds from subsites -2 to +2 in one monomer of the asymmetric unit, although it lies on subsites -1 to +3 in the other. These two "Michaelis complexes" have never been observed simultaneously before and are consistent with the dual mode of bond cleavage in this substrate. The bound sugar adopts a mixed 2₁ and 3₁ helical conformation similar to that reported in inactive mutants from families PL-1 and PL-10. However, our study suggests that the catalytic base in PelI is not a conventional arginine but a lysine as proposed in family PL-9.

Received for publication, December 5, 2007 , and in revised form, April 21, 2008.

The atomic coordinates and structure factors (code 3B4N, 3B90, and 3B8Y) 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 by grants from the Centre National de la Recherche Scientifique. 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 on-line version of this article (available at http://www.jbc.org) contains two supplemental tables.

¹ Present address: Division of Infectious Diseases, Children's Hospital, Harvard Medical School, Boston, MA 02115.

² Present address: Laboratoire Morphogenèse et Signalisation Cellulaire, UMR144 CNRS/Institut Curie, 26 Rue d'Ulm, 75248 Paris Cedex 05, France.

³ To whom correspondence should be addressed. Tel.: 33-472722624; Fax: 33-472722616; E-mail: p.gouet{at}ibcp.fr.

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