Protein targets of 1,4-benzoquinone and 1,4-naphthoquinone in human bronchial epithelial cells.
Many aspects of the toxicity of xenobiotic compounds have been attributed to the consequences of covalent modification of specific proteins, but the nature and specificity of protein targets for classes of electrophilic toxins remain largely uncharacterized. For inhaled toxicants, the point of exposure or absorption lies with epithelial cells lining the pulmonary tree. In this study, abundant proteins in human bronchial epithelial cells that are arylated in vitro by two quinonoid compounds, 1,4-benzoquinone (BQ) and 1,4-naphthoquinone (NQ) have been detected using (14)C-labeled quinones and two-dimensional gel electrophoresis. These proteins were identified using matrix assisted laser desorption/ionization mass spectrometry for tryptic mass mapping followed by sequence database searching. Corroborative identification of protein targets was obtained from the apparent isoelectric points, molecular weights, and the use of antibody probes. There were subtle differences in the protein targets of BQ and NQ, but both associated with the following abundant proteins, nucleophosmin, galectin-1, probable protein disulfide isomerase, protein disulfide isomerase, 60 kDa heat shock protein, mitochondrial stress-70 protein, epithelial cell marker protein, and S100-type calcium binding protein A14. We further delineate the properties of these proteins that make them preferred targets and the evidence these adducts present for delivery of these quinones to subcellular compartments.
Lamé
Michael W
MW
Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, CA 95616, USA.
Jones
A Daniel
AD
Wilson
Dennis W
DW
Segall
Hank J
HJ
D000293:Adolescent; D000328:Adult; D000595:Amino Acid Sequence; D001345:Autoradiography; D016227:Benzoquinones / Q000494:pharmacology*; D001980:Bronchi / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D004847:Epithelial Cells / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D009285:Naphthoquinones / Q000494:pharmacology*; D010084:Oxidation-Reduction; D010449:Peptide Mapping; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*
D016227:Benzoquinones; D009285:Naphthoquinones; D011506:Proteins; C004532:quinone; C035342:1,4-naphthoquinone