Identification of proteins adducted by reactive naphthalene metabolites in vitro

Margaret A. Isbell, Dexter Morin, Bridget Boland, Alan R Buckpitt, Michelle Salemi, Jack Presley

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Metabolic activation of inert chemicals to electrophilic intermediates has been correlated with the incidence and severity of cytotoxicity. The current studies have identified several proteins adducted by reactive metabolites of the lung toxicant, naphthalene. Proteins isolated from microsomal incubations of 14C-naphthalene were separated by 2-DE, proteins were blotted to PVDF membranes and radioactive proteins were localized by storage phosphor analysis. Adducted proteins were isolated from complimentary gels and identified by peptide mass mapping. A total of 18 adducted proteins were identified including: protein disulfide isomerase precursor, ER-60 protease, alpha actin, mouse urinary proteins, and cytochrome b5 reductase. In supernatant fractions, protein disulfide isomerase, heat shock protein 70, and α-actin were key proteins to which reactive naphthalene metabolites were bound. All of the proteins adducted, with the exception of cytochrome b5 reductase were sulfhydryl rich. Although several of the proteins found to be adducted in these studies have also been shown to be adducted by other electrophiles, several others have not been reported as common targets of reactive metabolites. These studies provide a basis for both in situ and in vivo work designed to follow the fate and formation of reactive metabolite protein adducts.

Original languageEnglish (US)
Pages (from-to)4197-4204
Number of pages8
JournalProteomics
Volume5
Issue number16
DOIs
StatePublished - Nov 2005

Fingerprint

Metabolites
Proteins
Cytochrome Reductases
Protein Disulfide-Isomerases
Cytochromes b5
Actins
Oxidoreductases
naphthalene
In Vitro Techniques
HSP70 Heat-Shock Proteins
Peptide Mapping
Cytotoxicity
Phosphors
Membrane Proteins
Gels
Chemical activation
Lung
Membranes
Peptides
Incidence

Keywords

  • Cytotoxicity
  • Electrophilic metabolites
  • Lung
  • Naphthalene
  • P450 monooxygenase

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Identification of proteins adducted by reactive naphthalene metabolites in vitro. / Isbell, Margaret A.; Morin, Dexter; Boland, Bridget; Buckpitt, Alan R; Salemi, Michelle; Presley, Jack.

In: Proteomics, Vol. 5, No. 16, 11.2005, p. 4197-4204.

Research output: Contribution to journalArticle

Isbell, MA, Morin, D, Boland, B, Buckpitt, AR, Salemi, M & Presley, J 2005, 'Identification of proteins adducted by reactive naphthalene metabolites in vitro', Proteomics, vol. 5, no. 16, pp. 4197-4204. https://doi.org/10.1002/pmic.200401278
Isbell, Margaret A. ; Morin, Dexter ; Boland, Bridget ; Buckpitt, Alan R ; Salemi, Michelle ; Presley, Jack. / Identification of proteins adducted by reactive naphthalene metabolites in vitro. In: Proteomics. 2005 ; Vol. 5, No. 16. pp. 4197-4204.
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