title: Selective enrichment and mass spectrometric identification of nitrated peptides using fluorinated carbon tags.
abstract: Protein tyrosine nitration (PTN) is a post-translational modification that is related to several acute or chronic diseases. PTN introduces a nitro group in the ortho position of the phenolic hydroxyl group of tyrosine residues. PTN has been shown to be involved in the pathogenesis of inflammatory responses, cancers, and neurodegenerative and age-related disorders. Furthermore, it has been proposed that PTN regulates signal cascades related to nitric oxide (NO·) production and NO-mediated processes. Although nitrated proteins as markers of oxidative stress are confirmed by immunological assays in various affected cells or tissues, it is not known how many different types of proteins in living cells are nitrated. Since protein nitration is a low-abundance post-translational modification, development of an effective enrichment method for nitrated proteins is needed to detect nitrated peptides or proteins from the limited amount of pathophysiological samples. In the present study, we developed an enrichment method using specific chemical tagging. Nitroproteome profiling using chemical tagging and mass spectrometry was validated by model proteins. Furthermore, we successfully identified numerous nitrated proteins from the Huh7 human hepatoma cell line.
authors.affiliation: Department of Molecular Biotechnology, WCU Program, Konkuk University School of Medicine, Seoul 143-701, Korea.
authors.forename: Jae Kyung
authors.initials: JK
authors.lastname: Kim
authors.forename: Jung Rok
authors.initials: JR
authors.lastname: Lee
authors.forename: Jeong Won
authors.initials: JW
authors.lastname: Kang
authors.forename: Soo Jae
authors.initials: SJ
authors.lastname: Lee
authors.forename: Gu Choul
authors.initials: GC
authors.lastname: Shin
authors.forename: Woon-Seok
authors.initials: WS
authors.lastname: Yeo
authors.forename: Kyun-Hwan
authors.initials: KH
authors.lastname: Kim
authors.forename: Hyung Soon
authors.initials: HS
authors.lastname: Park
authors.forename: Kwang Pyo
authors.initials: KP
authors.lastname: Kim
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002244:Carbon / Q000737:chemistry*; D002417:Cattle; D002457:Cell Extracts; D045744:Cell Line, Tumor; D019295:Computational Biology; D005240:Feasibility Studies; D005461:Fluorine / Q000737:chemistry; D054879:Halogenation; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D009574:Nitro Compounds / Q000737:chemistry*; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification; D010455:Peptides / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism; D052616:Solid Phase Extraction; D014357:Trypsin / Q000378:metabolism; D014443:Tyrosine / Q000737:chemistry
chemical_list: D002457:Cell Extracts; D009574:Nitro Compounds; D010446:Peptide Fragments; D010455:Peptides; D012710:Serum Albumin, Bovine; D005461:Fluorine; D014443:Tyrosine; D002244:Carbon; D014357:Trypsin