Reactive naphthalene metabolite binding to hemoglobin and albumin.
Earlier work has shown that the murine Clara cell cytotoxicant, naphthalene, is metabolized to reactive metabolites which deplete glutathione or, in the absence of sufficient glutathione, become bound covalently to tissue macromolecules. Correlations between bound metabolite levels in the lung with injury suggests an association between reactive metabolite binding and toxicity. In this study we examine the formation of covalent naphthalene adducts with hemoglobin and albumin in mice to determine whether these serve as useful indices of exposure and metabolism for a chemical which shows a glutathione threshold. Covalent binding of radioactivity from [3H]naphthalene to both albumin and hemoglobin was dose dependent and a glutathione threshold was observed. At early times after naphthalene administration, the formation of albumin adducts was 10- to 30-fold higher than that of hemoglobin adducts. Hemoglobin and albumin adduct levels decreased by apparent first-order processes with half-lives of 11.5 and 1.8 days, respectively. These half-lives are consistent with the turnover of these blood proteins in the mouse. Pretreatment with buthionine sulfoximine resulted in higher levels of albumin adduct but in no alteration of hemoglobin adduct levels in comparison with control. In contrast, diethylmaleate pretreatment increased the level of hemoglobin adduct but not albumin adduct. The antibody to naphthalene mercapturates recognized the hemoglobin adduct(s) but not the albumin adduct(s). Comparison of the data from ELISA (standardized using hydroxymercaptodihydronaphthalene) and radiochemical analysis yielded curves with identical slopes; the absolute levels of adduct found by ELISA were approximately half those measured with radiochemical techniques.(ABSTRACT TRUNCATED AT 250 WORDS)
Cho
M
M
Department of Pharmacology and Toxicology, School of Veterinary Medicine, University of California at Davis 95616.
Jedrychowski
R
R
Hammock
B
B
Buckpitt
A
A
D000818:Animals; D019328:Buthionine Sulfoximine; D004591:Electrophoresis, Polyacrylamide Gel; D005978:Glutathione / Q000037:antagonists & inhibitors / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism*; D008297:Male; D008298:Maleates / Q000494:pharmacology; D008717:Methionine Sulfoximine / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D009281:Naphthalenes / Q000378:metabolism*; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism
D006454:Hemoglobins; D008298:Maleates; D009281:Naphthalenes; D012709:Serum Albumin; D013438:Sulfhydryl Compounds; D008717:Methionine Sulfoximine; C031721:naphthalene; D019328:Buthionine Sulfoximine; C014476:diethyl maleate; D005978:Glutathione