Titre :

The structural, dynamical and biochemical characterizations ofVerticillium dahliaepectate 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]
Molinié, Roland [Auteur]
Fontaine, Jean-Xavier [Auteur]
Institut Charles Viollette (ICV) - ULR 7394
Lemaire, Adrien [Auteur]
Voxeur, Aline [Auteur]
Pilard, Serge [Auteur]
Pau-Roblot, Corinne [Auteur]
Mercadante, Davide [Auteur]
Pelloux, Jérôme [Auteur]
Sénéchal, Fabien [Auteur]

Titre de la revue :

BioRxiv

Éditeur :

Cold Spring Harbor Laboratory

Date de publication :

2022-11-09

Discipline(s) HAL :

Sciences du Vivant [q-bio]

Résumé en anglais : [en]

AbstractPectins, 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 >AbstractPectins, 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 fromVerticillium 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 fromVerticillium-partially tolerant and sensitive flax cultivars differed which could facilitate the identification VdPelB-mediated elicitors of defence responses.Lire moins >

Langue :

Anglais

Audience :

Non spécifiée

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