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S-arylcysteine-keratin adducts as biomarkers of human dermal exposure to aromatic hydrocarbons

Title
S-arylcysteine-keratin adducts as biomarkers of human dermal exposure to aromatic hydrocarbons
Author
남태규
Keywords
MURINE CLARA CELLS; JET FUEL JP-8; STRATUM-CORNEUM; BENZENE METABOLISM; CYTOCHROMES P450; PROTEIN ADDUCTS; ARENE OXIDES; NAPHTHALENE; HEMOGLOBIN; EXPRESSION
Issue Date
2008-04
Publisher
AMER CHEMICAL SOC
Citation
CHEMICAL RESEARCH IN TOXICOLOGY, v. 21, No. 4, Page. 852-858
Abstract
To measure biomarkers of skin exposure to ubiquitous industrial and environmental aromatic hydrocarbons, we sought to develop an ELISA to quantitate protein adducts of metabolites of benzene and naphthalene in the skin of exposed individuals. We hypothesized that electrophilic arene oxides formed by CYP isoforms expressed in the human skin react with nucleophilic sites on keratin, the most abundant protein in the stratum corneum that is synthesized de novo during keratinocyte maturation and differentiation. The sulthydryl groups of cysteines in the head region of the keratin proteins 1 (K1) and 10 (K10) are likely targets. The following synthetic S-arylcysteines were incorporated into 10-mer head sequences of K1 [GGGRFSS(S-aryl-QGG] and K10 [GGGG(S-aryl-C)GGGGG] to form the predicted immunogenic epitopes for antibody production for ELISA: S-phenylcysteine-K1 (SPK1), S-phenyleysteine-K10 (SPK10), S-(1-naphthyl)cysteine-K1 (1NK1), S-(1-naphthyl)cysteine-K10 (1NK10), S-(2-naphthyl)cysteine-K1 (2NK1), and S-(2-naphthyl)cysteine-K10 (2NK 10). Analysis by ELISA was chosen based on its high throughput and sensitivity, and low cost. The synthetic modified oligopeptides, available in quantity, served both as immunogens and as chemical standards for quantitative ELISA. Polyclonal rabbit antibodies produced against the naphthyl-modified keratins reacted with their respective antigens with threshold sensitivities of 15-31 ng/mL and high specificity over a linear range up to 500 ng/mL. Anti-S-phenylcysteine antibodies were not sufficiently specific or sensitive toward the target antigens for use in ELISA under our experimental conditions. In dermal tape-strip samples collected from 13 individuals exposed to naphthalene-containing jet fuel, naphthyl-conjugated peptides were detected at levels from 0.343 +/- 0.274 to 2.34 +/- 1.61 pmol adduct/mu g keratin but were undetectable in unexposed volunteers. This is the first report of adducts of naphthalene (or of any polycyclic aromatic hydrocarbon) detected in the exposed intact human skin. Quantitation of naphthyl-keratin adducts in the skin of exposed individuals will allow us to investigate the importance of dermal penetration, metabolism, and adduction to keratin and to predict more accurately the contribution of dermal exposure to systemic dose for use in exposure and risk-assessment models.
URI
https://pubs.acs.org/doi/abs/10.1021/tx7003773http://repository.hanyang.ac.kr/handle/20.500.11754/104454
ISSN
0893-228X
DOI
10.1021/tx7003773
Appears in Collections:
COLLEGE OF PHARMACY[E](약학대학) > PHARMACY(약학과) > Articles
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