title: Proteomic analysis of S-nitrosylation induced by 1-methyl-4-phenylpyridinium (MPP+).
abstract: BACKGROUND
  Nitric oxide (NO) mediates its function through the direct modification of various cellular targets. S-nitrosylation is a post-translational modification of cysteine residues by NO that regulates protein function. Recently, an imbalance of S-nitrosylation has also been linked to neurodegeneration through the impairment of pro-survival proteins by S-nitrosylation.
  
  
  RESULTS
  In the present study, we used two-dimensional gel electrophoresis in conjunction with the modified biotin switch assay for protein S-nitrosothiols using resin-assisted capture (SNO-RAC) to identify proteins that are S-nitrosylated more intensively in neuroblastoma cells treated with a mitochondrial complex I inhibitor, 1-methyl-4-phenylpyridinium (MPP+). We identified 14 proteins for which S-nitrosylation was upregulated and seven proteins for which it was downregulated in MPP+-treated neuroblastoma cells. Immunoblot analysis following SNO-RAC confirmed a large increase in the S-nitrosylation of esterase D (ESD), serine-threonine kinase receptor-associated protein (STRAP) and T-complex protein 1 subunit γ (TCP-1 γ) in MPP+-treated neuroblastoma cells, whereas S-nitrosylation of thioredoxin domain-containing protein 5 precursor (ERp46) was decreased.
  
  
  CONCLUSIONS
  These results suggest that S-nitrosylation resulting from mitochondrial dysfunction can compromise neuronal survival through altering multiple signal transduction pathways and might be a potential therapeutic target for neurodegenerative diseases.
authors.affiliation: Department of Pharmacology, Nara Medical University School of Medicine, 840 Shijo-cho, Kashihara, Nar, 634-8521, Japan.|Department of Genomic Drug Discovery Science, Kyoto University Graduate School of Pharmaceutical Sciences, Sakyo-ku, Kyoto, Japan.
authors.forename: Akira T
authors.initials: AT
authors.lastname: Komatsubara
authors.affiliation: Division of Functional Genomics, Advanced Science Research Center, Kanazawa University, Kanazawa, Japan.
authors.forename: Tomoya
authors.initials: T
authors.lastname: Asano
authors.affiliation: World-Leading Drug Discovery Research Center, Kyoto University, Sakyo-ku, Kyoto, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Tsumoto
authors.affiliation: World-Leading Drug Discovery Research Center, Kyoto University, Sakyo-ku, Kyoto, Japan.|Department of Nanobio Drug Discovery, Kyoto University Graduate School of Pharmaceutical Sciences, Sakyo-ku, Kyoto, 606-8501, Japan.
authors.forename: Kazuharu
authors.initials: K
authors.lastname: Shimizu
authors.affiliation: Division of Functional Genomics, Advanced Science Research Center, Kanazawa University, Kanazawa, Japan.
authors.forename: Takumi
authors.initials: T
authors.lastname: Nishiuchi
authors.affiliation: Department of Pharmacology, Nara Medical University School of Medicine, 840 Shijo-cho, Kashihara, Nar, 634-8521, Japan.
authors.forename: Masanori
authors.initials: M
authors.lastname: Yoshizumi
authors.affiliation: Department of Pharmacology, Nara Medical University School of Medicine, 840 Shijo-cho, Kashihara, Nar, 634-8521, Japan.|World-Leading Drug Discovery Research Center, Kyoto University, Sakyo-ku, Kyoto, Japan.
authors.forename: Kentaro
authors.initials: K
authors.lastname: Ozawa