Titre :

The structural, dynamical and biochemical characterizations of Verticillium dahliae pectate lyase, VdPelB, highlight its specificities

Auteur(s) :

Safran, Josip [Auteur]
Ung, Vanessa [Auteur]
Bouckaert, Julie [Auteur]
Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576
Habrylo, Olivier [Auteur]
Molinie, Roland [Auteur]
BioEcoAgro - UMR transfrontalière INRAe - UMRT1158
Institut Charles Viollette (ICV) - ULR 7394
Fontaine, Jean-Xavier [Auteur]
BioEcoAgro - UMR transfrontalière INRAe - UMRT1158
Institut Charles Viollette (ICV) - ULR 7394
Lemaire, Adrien [Auteur]
Voxeur, Aline [Auteur]
Pilard, Serge [Auteur]
Pau, Corinne [Auteur]
Institut Charles Viollette (ICV) - ULR 7394
BioEcoAgro - UMR transfrontalière INRAe - UMRT1158
Mercadante, Davide [Auteur]
Pelloux, Jerome [Auteur]
BioEcoAgro - UMR transfrontalière INRAe - UMRT1158
Institut Charles Viollette (ICV) - ULR 7394
Senechal, Fabien [Auteur]
Institut Charles Viollette (ICV) - ULR 7394
BioEcoAgro - UMR transfrontalière INRAe - UMRT1158

Titre de la revue :

BioRxiv

Éditeur :

Cold Spring Harbor Laboratory

Date de publication :

2022-11-09

Mot(s)-clé(s) en anglais :

Pectate lyase
pectins
homogalacturonan
oligogalacturonides
Flax
Verticillium dahliae

Discipline(s) HAL :

Sciences du Vivant [q-bio]
Chimie/Chimie théorique et/ou physique
Sciences de l'environnement/Biodiversité et Ecologie
Sciences du Vivant [q-bio]/Biochimie, Biologie Moléculaire/Biochimie [q-bio.BM]
Sciences du Vivant [q-bio]/Biochimie, Biologie Moléculaire/Biologie structurale [q-bio.BM]
Sciences du Vivant [q-bio]/Biotechnologies

Résumé en anglais : [en]

Abstract Pectins, complex polysaccharides and major components of the plant primary cell wall, can be degraded by pectate lyases (PLs). PLs cleave glycosidic bonds of homogalacturonans (HG), the main pectic domain, by ...
Lire la suite >Abstract Pectins, complex polysaccharides and major components of the plant primary cell wall, can be degraded by pectate lyases (PLs). PLs cleave glycosidic bonds of homogalacturonans (HG), the main pectic domain, by β-elimination, releasing unsaturated oligogalacturonides (OGs). To understand the catalytic mechanism and structure/function of these enzymes, we characterized VdPelB from Verticillium dahliae, a plant pathogen. We first solved the crystal structure of VdPelB at 1.2Å resolution showing that it is a right-handed parallel β-helix structure. Molecular dynamics (MD) simulations further highlighted the dynamics of the enzyme in complex with substrates that vary in their degree of methylesterification, identifying amino acids involved in substrate binding and cleavage of non-methylesterified pectins. We then biochemically characterized wild type and mutated forms of VdPelB. VdPelB was most active on non-methylesterified pectins, at pH 8 in presence of Ca2+ions. VdPelB-G125R mutant was most active at pH 9 and showed higher relative activity compared to native enzyme. The OGs released by VdPelB differed to that of previously characterized PLs, showing its peculiar specificity in relation to its structure. OGs released from Verticillium-partially tolerant and sensitive flax cultivars differed which could facilitate the identification VdPelB-mediated elicitors of defence responses.Lire moins >

Langue :

Anglais

Comité de lecture :

Non

Audience :

Internationale

Vulgarisation :

Non

Établissement(s) :

Université de Lille
CNRS

Collections :

• BioEcoAgro - UMR-T 1158
• Unité de Glycobiologie Structurale et Fonctionnelle (UGSF) - UMR 8576

Équipe(s) de recherche :

Computational Molecular Systems Biology

Date de dépôt :

2022-12-19T15:41:25Z
2023-01-04T08:19:29Z
2023-01-04T11:21:46Z
2023-01-04T11:57:13Z