title: Copper stress-induced changes in leaf soluble proteome of Cu-sensitive and tolerant Agrostis capillaris L. populations.
abstract: Changes in leaf soluble proteome were explored in 3-month-old plants of metallicolous (M) and nonmetallicolous (NM) Agrostis capillaris L. populations exposed to increasing Cu concentrations (1-50 μM) to investigate molecular mechanisms underlying plant responses to Cu excess and tolerance of M plants. Plants were cultivated on perlite (CuSO4 spiked-nutrient solution). Soluble proteins, extracted by the trichloroacetic acid/acetone procedure, were separated with 2-DE (linear 4-7 pH gradient). Analysis of CCB-stained gels (PDQuest) reproducibly detected 214 spots, and 64 proteins differentially expressed were identified using LC-MS/MS. In both populations, Cu excess impacted both light-dependent (OEE, cytochrome b6-f complex, and chlorophyll a-b binding protein), and -independent (RuBisCO) photosynthesis reactions, more intensively in NM leaves (ferredoxin-NADP reductase and metalloprotease FTSH2). In both populations, upregulation of isocitrate dehydrogenase and cysteine/methionine synthases respectively suggested increased isocitrate oxidation and enhanced need for S-containing amino-acids, likely for chelation and detoxification. In NM leaves, an increasing need for energetic compounds was indicated by the stimulation of ATPases, glycolysis, pentose phosphate pathway, and Calvin cycle enzymes; impacts on protein metabolism and oxidative stress increase were respectively suggested by the rise of chaperones and redox enzymes. Overexpression of a HSP70 may be pivotal for M Cu tolerance by protecting protein metabolism. All MS data have been deposited in the ProteomeXchange with the dataset identifier PXD001930 (http//proteomecentral.proteomexchange.org/dataset/PXD001930).
authors.affiliation: UMR1202 BIOGECO, University of Bordeaux, Pessac Cedex, France.|UMR1202 BIOGECO, INRA, Cestas cedex, France.
authors.forename: Elena
authors.initials: E
authors.lastname: Hego
authors.affiliation: Bordeaux INP, Univ. Bordeaux, Bordeaux, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Vilain
authors.affiliation: Centre de Génomique Fonctionnelle, Centre de Bioinformatique de Bordeaux, Univ. Bordeaux, Bordeaux, France.
authors.forename: Aurélien
authors.initials: A
authors.lastname: Barré
authors.affiliation: Centre de Génomique Fonctionnelle, Plateforme Protéome, Univ. Bordeaux, Bordeaux, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Claverol
authors.affiliation: Centre de Génomique Fonctionnelle, Plateforme Protéome, Univ. Bordeaux, Bordeaux, France.
authors.forename: Jean-William
authors.initials: JW
authors.lastname: Dupuy
authors.affiliation: UMR1202 BIOGECO, University of Bordeaux, Pessac Cedex, France.|UMR1202 BIOGECO, INRA, Cestas cedex, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Lalanne
authors.affiliation: Bordeaux INP, Centre de Génomique Fonctionnelle, Plateforme Protéome, Univ. Bordeaux, France.
authors.forename: Marc
authors.initials: M
authors.lastname: Bonneu
authors.affiliation: UMR1202 BIOGECO, University of Bordeaux, Pessac Cedex, France.|UMR1202 BIOGECO, INRA, Cestas cedex, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Plomion
authors.affiliation: UMR1202 BIOGECO, University of Bordeaux, Pessac Cedex, France.|UMR1202 BIOGECO, INRA, Cestas cedex, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Mench
mesh_terms: D000222:Adaptation, Physiological / Q000235:genetics*; D031703:Agrostis / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D002734:Chlorophyll / Q000235:genetics / Q000378:metabolism; D000077194:Chlorophyll A; D061146:Chlorophyll Binding Proteins / Q000235:genetics / Q000378:metabolism; D019327:Copper Sulfate / Q000633:toxicity*; D045346:Cytochrome b6f Complex / Q000235:genetics / Q000378:metabolism; D004734:Energy Metabolism / Q000187:drug effects / Q000235:genetics; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D063990:Gene Ontology; D058977:Molecular Sequence Annotation; D010788:Photosynthesis / Q000187:drug effects / Q000235:genetics; D018515:Plant Leaves / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D020543:Proteome / Q000235:genetics* / Q000378:metabolism; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics / Q000378:metabolism; D012995:Solubility; D013312:Stress, Physiological
chemical_list: D061146:Chlorophyll Binding Proteins; D020543:Proteome; D002734:Chlorophyll; C037184:chlorophyll b; D045346:Cytochrome b6f Complex; D012273:Ribulose-Bisphosphate Carboxylase; D019327:Copper Sulfate; D000077194:Chlorophyll A
keywords: Heat-shock protein; MS-LC-MS/MS; Oxidative stress; Photosynthesis; Plant proteomics