Literature DB >> 20972763

One-dimensional western blotting coupled to LC-MS/MS analysis to identify chemical-adducted proteins in rat urine.

Matthew T Labenski1, Ashley A Fisher, Terrence J Monks, Serrine S Lau.   

Abstract

The environmental toxicant hydroquinone (HQ) and its glutathione conjugates (GSHQs) cause renal cell necrosis via a combination of redox cycling and the covalent adduction of proteins within the S₃ segment of the renal proximal tubules in the outer stripe of the outer medulla (OSOM). Following administration of 2-(glutathion-S-yl)HQ (MGHQ) (400 μmol/kg, i.v., 2 h) to Long Evans (wild-type Eker) rats, Western analysis utilizing an antibody specific for quinol-thioether metabolites of HQ revealed the presence of large amounts of chemical-protein adducts in both the OSOM and urine. By aligning the Western blot film with a parallel gel stained for protein, we can isolate the adducted proteins for LC-MS/MS analysis. Subsequent database searching can identify the specific site(s) of chemical adduction within these proteins. Finally, a combination of software programs can validate the identity of the adducted peptides. The site-specific identification of covalently adducted and oxidized proteins is a prerequisite for understanding the biological significance of chemical-induced posttranslational modifications (PTMs) and their toxicological significance.

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Year:  2011        PMID: 20972763      PMCID: PMC4096012          DOI: 10.1007/978-1-60761-849-2_20

Source DB:  PubMed          Journal:  Methods Mol Biol        ISSN: 1064-3745


  8 in total

1.  Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels.

Authors:  A Shevchenko; M Wilm; O Vorm; M Mann
Journal:  Anal Chem       Date:  1996-03-01       Impact factor: 6.986

2.  Method to correlate tandem mass spectra of modified peptides to amino acid sequences in the protein database.

Authors:  J R Yates; J K Eng; A L McCormack; D Schieltz
Journal:  Anal Chem       Date:  1995-04-15       Impact factor: 6.986

3.  Carcinogenicity of a nephrotoxic metabolite of the "nongenotoxic" carcinogen hydroquinone.

Authors:  S S Lau; T J Monks; J I Everitt; E Kleymenova; C L Walker
Journal:  Chem Res Toxicol       Date:  2001-01       Impact factor: 3.739

4.  Investigative proteomics: identification of an unknown plant virus from infected plants using mass spectrometry.

Authors:  Bret Cooper; Donna Eckert; Nancy L Andon; John R Yates; Paul A Haynes
Journal:  J Am Soc Mass Spectrom       Date:  2003-07       Impact factor: 3.109

5.  Quinone electrophiles selectively adduct "electrophile binding motifs" within cytochrome c.

Authors:  Ashley A Fisher; Matthew T Labenski; Srinivas Malladi; Vijay Gokhale; Martina E Bowen; Rania S Milleron; Shawn B Bratton; Terrence J Monks; Serrine S Lau
Journal:  Biochemistry       Date:  2007-09-07       Impact factor: 3.162

6.  Proteomic characterization of wheat amyloplasts using identification of proteins by tandem mass spectrometry.

Authors:  Nancy L Andon; Sarah Hollingworth; Antonius Koller; Andrew J Greenland; John R Yates; Paul A Haynes
Journal:  Proteomics       Date:  2002-09       Impact factor: 3.984

7.  Immunochemical analysis of quinol-thioether-derived covalent protein adducts in rodent species sensitive and resistant to quinol-thioether-mediated nephrotoxicity.

Authors:  H E Kleiner; T W Jones; T J Monks; S S Lau
Journal:  Chem Res Toxicol       Date:  1998-11       Impact factor: 3.739

8.  Immunochemical detection of quinol--thioether-derived protein adducts.

Authors:  H E Kleiner; M I Rivera; N R Pumford; T J Monks; S S Lau
Journal:  Chem Res Toxicol       Date:  1998-11       Impact factor: 3.739

  8 in total
  1 in total

1.  Mapping Adverse Outcome Pathways for Kidney Injury as a Basis for the Development of Mechanism-Based Animal-Sparing Approaches to Assessment of Nephrotoxicity.

Authors:  Angela Mally; Sebastian Jarzina
Journal:  Front Toxicol       Date:  2022-06-15
  1 in total

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