title: S-nitroso-proteome in poplar leaves in response to acute ozone stress.
abstract: Protein S-nitrosylation, the covalent binding of nitric oxide (NO) to protein cysteine residues, is one of the main mechanisms of NO signaling in plant and animal cells. Using a combination of the biotin switch assay and label-free LC-MS/MS analysis, we revealed the S-nitroso-proteome of the woody model plant Populus x canescens. Under normal conditions, constitutively S-nitrosylated proteins in poplar leaves and calli comprise all aspects of primary and secondary metabolism. Acute ozone fumigation was applied to elicit ROS-mediated changes of the S-nitroso-proteome. This treatment changed the total nitrite and nitrosothiol contents of poplar leaves and affected the homeostasis of 32 S-nitrosylated proteins. Multivariate data analysis revealed that ozone exposure negatively affected the S-nitrosylation status of leaf proteins: 23 proteins were de-nitrosylated and 9 proteins had increased S-nitrosylation content compared to the control. Phenylalanine ammonia-lyase 2 (log2[ozone/control] = -3.6) and caffeic acid O-methyltransferase (-3.4), key enzymes catalyzing important steps in the phenylpropanoid and subsequent lignin biosynthetic pathways, respectively, were de-nitrosylated upon ozone stress. Measuring the in vivo and in vitro phenylalanine ammonia-lyase activity indicated that the increase of the phenylalanine ammonia-lyase activity in response to acute ozone is partly regulated by de-nitrosylation, which might favor a higher metabolic flux through the phenylpropanoid pathway within minutes after ozone exposure.
authors.affiliation: Research Unit Environmental Simulation, Institute for Biochemical Plant Pathology, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Elisa
authors.initials: E
authors.lastname: Vanzo
authors.affiliation: Research Unit Environmental Simulation, Institute for Biochemical Plant Pathology, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Ghirardo
authors.affiliation: Research Unit Protein Science, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Merl-Pham
authors.affiliation: Institute for Biochemical Plant Pathology, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Lindermayr
authors.affiliation: Institute for Biochemical Plant Pathology, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Werner
authors.initials: W
authors.lastname: Heller
authors.affiliation: Research Unit Protein Science, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Stefanie M
authors.initials: SM
authors.lastname: Hauck
authors.affiliation: Institute for Biochemical Plant Pathology, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Durner
authors.affiliation: Research Unit Environmental Simulation, Institute for Biochemical Plant Pathology, Helmholtz Zentrum München, Neuherberg, Germany.
authors.forename: Jörg-Peter
authors.initials: JP
authors.lastname: Schnitzler
mesh_terms: D053858:Metabolic Networks and Pathways / Q000187:drug effects; D008780:Methyltransferases / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism / Q000494:pharmacology; D009603:Nitroso Compounds / Q000032:analysis / Q000378:metabolism*; D010126:Ozone / Q000494:pharmacology*; D010650:Phenylalanine Ammonia-Lyase / Q000378:metabolism; D018515:Plant Leaves / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D010940:Plant Proteins / Q000032:analysis / Q000378:metabolism*; D032107:Populus / Q000187:drug effects* / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D020543:Proteome / Q000187:drug effects* / Q000378:metabolism; D026403:S-Nitrosothiols / Q000032:analysis / Q000737:chemistry; D013312:Stress, Physiological / Q000187:drug effects; D013997:Time Factors
chemical_list: D009603:Nitroso Compounds; D010940:Plant Proteins; D020543:Proteome; D026403:S-Nitrosothiols; D009569:Nitric Oxide; D010126:Ozone; D008780:Methyltransferases; C019608:caffeate O-methyltransferase; D010650:Phenylalanine Ammonia-Lyase