Substrate specificity of plant and fungi pectin methylesterases: Identification of novel inhibitors of PMEs.
Pectin methylesterases (PMEs) play a central role in pectin remodeling during plant development. They are also present in phytopathogens such as bacteria and fungi. We investigated the substrate specificity and pH dependence of plant and fungi PMEs using tailor-made pectic substrates. For this purpose, we used two plant PMEs (from orange peel: Citrus sinensis and from Arabidopsis thaliana) and one fungal PME (from Botrytis cinerea). We showed that plant and fungi PMEs differed in their substrate specificity and pH dependence, and that there were some differences between plant PMEs. We further investigated the inhibition of these enzyme activities using characterized polyphenols such as catechins and tannic acid. We showed that PMEs differed in their sensitivity to chemical compounds. In particular, fungal PME was not sensitive to inhibition. Finally, we screened for novel chemical inhibitors of PMEs using a chemical library of ∼3600 compounds. We identified a hundred new inhibitors of plant PMEs, but none had an effect on the fungal enzyme. This study sheds new light on the specificity of pectin methylesterases and provides new tools to modulate their activity.
Unité de Biologie des Plantes et Innovation, EA3900, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France.
Mélanie
M
L'Enfant
Unité de Biologie des Plantes et Innovation, EA3900, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France.
Jean-Marc
JM
Domon
Unité de Biologie des Plantes et Innovation, EA3900, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France.
Catherine
C
Rayon
CEA Cadarache, SBVME/LBDP, Bât 178, 13108 Saint-Paul-les-Durance Cedex, France.
Thierry
T
Desnos
INRA, UR 1268, Biopolymères, Interactions, Assemblages, BP 71627, 44316 Nantes Cedex 03, France.
Marie-Christine
MC
Ralet
INRA, UR 1268, Biopolymères, Interactions, Assemblages, BP 71627, 44316 Nantes Cedex 03, France.
Estelle
E
Bonnin
Unité de Biologie des Plantes et Innovation, EA3900, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France.
Jérôme
J
Pelloux
Unité de Biologie des Plantes et Innovation, EA3900, Université de Picardie Jules Verne, 33 rue Saint Leu, 80039 Amiens Cedex, France. Electronic address: corinne.pau-roblot@u-picardie.fr.
Corinne
C
Pau-Roblot
D000595:Amino Acid Sequence; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D055808:Drug Discovery; D004353:Drug Evaluation, Preclinical; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology; D005658:Fungi / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010944:Plants / Q000201:enzymology*; D059808:Polyphenols / Q000494:pharmacology; D016415:Sequence Alignment; D054852:Small Molecule Libraries; D013379:Substrate Specificity
D004791:Enzyme Inhibitors; D059808:Polyphenols; D054852:Small Molecule Libraries; D002265:Carboxylic Ester Hydrolases; C022323:pectinesterase
Catechins; Chemical inhibitors; Chemical library; EGCG; Pectin methylesterase