Identification of proteins adducted by reactive naphthalene metabolites in vitro
| Autor: | Alan R. Buckpitt, Jack M. Presley, Margaret A. Isbell, Bridget C. Boland, Dexter Morin, Michelle Salemi |
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| Rok vydání: | 2005 |
| Předmět: |
Proteome
Metabolite medicine.medical_treatment Molecular Sequence Data Naphthalenes Proteomics Biochemistry Peptide Mapping Mixed Function Oxygenases chemistry.chemical_compound Mice medicine Animals Electrophoresis Gel Two-Dimensional Amino Acid Sequence Carbon Radioisotopes Protein disulfide-isomerase Molecular Biology Cytochrome b5 reductase Protease Chemistry In vitro Hsp70 Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Microsome Microsomes Liver |
| Zdroj: | Proteomics. 5(16) |
| ISSN: | 1615-9853 |
| Popis: | 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 (14)C-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 alpha-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. |
| Databáze: | OpenAIRE |
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