Abstract
Incubation of 14C-labeled 3,4,3′,4′-tetrachlorobiphenyl (TCB) with a reconstituted monooxygenase system containing cytochrome P-450 isolated from 3-methylcholanthrene (MC)-treated rats caused an increase in incorporation of radioactivity to added rat erythrocytes and resulted in covalent binding of reactive metabolites to hemoglobin. Both increases in incorporation and covalent binding to the proteins were dependent on incubation time and required cytochrome P-450-linked monooxygenase activity. In the absence of erythrocytes, the reactive TCB metabolites bound to cytochrome P-450, NADPH-cytochrome P-450 reductase, and high-molecular weight aggregates, as determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, whereas inclusion of the cells in the reaction mixture caused marked alkylation of hemoglobin and a decrease in binding to the electron transfer proteins. Several metabolites other than protein-bound metabolites were detected by thin-layer chromatography; some were found to be incorporated into the erythrocytes while others were not. These results suggest that several TCB metabolites formed by cytochrome P-450 can interact with rat erythrocytes in vitro, and that these interactions may be responsible for the persistence of TCB metabolites in blood observed in vivo.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abdel-Hamid FM, Moore LA, Matthews HB (1981) Comparative study of 3,4,3′,4′-tetrachlorobiphenyl in male and female rats and female monkeys. J Toxicol Environ Health 7: 181–191
Albrecht G, Kiese M, Sies H, Weger N (1976) The effect of Tris(2-chloroethyl)amine on human hemoglobin. Arch Pharmacol 294: 179–185
Allen JR, Carstens LA, Barsotti DA (1974) Residual effects of short-term, low level exposure of nonhuman primates to polychlorinated biphenyls. Toxicol Appl Pharmacol 30: 440–451
Bandiere S, Safe S, Okey AB (1982) Binding of polychlorinated biphenyls classified as either phenobarbitone, 3-methylcholanthrene, or mixed-type inducers to cytosolic Ah receptor. Chem Biol Interact 39: 259–277
Fujita K, Iwasaki T, Mori (1979) Serum inhibitor active against hemolytic activity of polychlorinated biphenyls. Fukuoka Acta Med 70: 99–101
Goldstein J (1980) Structure-activity relationships for the biochemical effects and the relationship to toxicity. In Kimbrough (ed), Halogenated biphenyls, terphenyls, naphthalenes, dibenzodioxins and related products. Elsevier/North-Holland Biomedical Press, pp151–190
Harada N, Omura T (1981) Selective induction of two different molecular species of cytochrome P-450 by phenobarbital and 3-methylcholanthrene. J Biochem 89: 237–248
Hori S, Obana H, Kashimoto T, Otake T, Nishimura H, Ikegami N, Kunita N, Fukuda Y, Uda H (1982) Biological effects of the compounds related to Yusho on female cynomolgus monkeys (Macaca fascicularis). II. Studies on biochemical finding of blood, hepatic microsomal enzyme, immune response and histopathology. Proc Osaka Pref Inst Public Health 12: 9–25
Imai Y (1981) The use of 8-aminooctyl Sepharose for the separation of some components of the hepatic microsomal electron transfer system. J Biochem 80: 267–276
Kim S, Paik WK, Choi J, Lotlikar PD, Magee PN (1981) Microsome-dependent methylation of erythrocyte proteins by dimethylnitrosoamine. Carcinogenesis 2: 179–182
Kuratsune M (1972) An abstract of results of laboratory examinations of patients with Yusho and of animal experiments. Environ Health Perspect 1: 129–136
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature (London) 227: 680–685
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275
Lutz RJ, Dedrick RL, Matthews HB, Eling TE, Anderson MW (1977) A preliminary pharmacokinetic model for several chlorinated biphenyls in the rat. Drug Metab Dispos 5: 386–396
Matsumura F (1983) Biochemical aspects of action mechanism of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related chemicals in animals. Pharmacol Ther 19: 195–209
Matthews HB, Anderson MW (1975) Effects of chlorination on the distribution and excretion of polychlorinated biphenyls. Drug Metab Dispos 3: 371–380
Miller TL (1978) Interaction of hydroxychlorobiphenyl-polychlorinated biphenyl metabolites with the human erythrocyte membrane. J Environ Path Toxicol 1: 459–474
Mori R, Fujita K, Yamaguchi M, Yoshimura H, Yamamoto H (1975) Effect of polychlorinated biphenyl (PCB) on HeLa cells. Fukuoka Acta Med 66: 568–573
Neuman HG (1984) Analysis of hemoglobin as a dose monitor for alkylating and arylating agents. Arch Toxicol 56: 1–6
Omura T, Sato R (1964) The carbon monoxide binding pigment of liver microsomes. J Biol Chem 239: 2379–2385
Osterman-Golkar S, Ehrenberg L, Segerback D, Hallstrom I (1976) Evaluation of genetic risks of alkylating agents. II. hemoglobin as a dose monitor. Mutat Res 34: 1–10
Parkinson A, Safe S (1981) Aryl hydrocarbon hydroxylase induction and its relationship to the toxicity of halogenated aryl hydrocarbons. Toxicol Environ Chem Rev 4: 1–46
Perreira MA, Chang LW (1981) Binding of chemical carcinogens and mutagens to rat hemoglobin. Chem Biol Interact 33: 301–305
Perreira MA, Chang LW (1982) Binding of chlorofolm to mouse and rat hemoglobin. Chem Biol Interact 39: 89–99
Poland A, Glover E (1977) Chlorinated biphenyl induction of aryl hydrocarbon hydroxylase activity: a study of the structure-activity relationship. Mol Pharmacol 13: 924–938
Poland A, Knutson JC (1982) 2,3,7,8-Tetrachlorodibenzo-p-dioxin and related halogenated aromatic hydrocarbons: examination of the mechanism of toxicity. Ann Rev Pharmacol Toxicol 22: 517–554
Safe S (1980) Metabolism, uptake, storage and bioaccumulation. In: Kimbrough RD (ed) Halogenated biphenyls, terphenyls, naphthalenes, dibenzodioxins and related products. Elsevier/ North-Holland Biomedical Press, pp 81–107
Segerback D (1983) Alkylation of DNA and hemoglobin in the mouse following exposure to ethene and ethene oxide. Chem Biol Interact 45: 139–151
Shimada T, Nunoura Y, Kitanaka E, Iwagami S, Mizuta Y (1977) Recovery from toxicological and biochemical effects of rats intoxicated with polychlorinated biphenyls. Folia Pharmacol Jpn 73: 887–894
Shimada T, Sato R (1980) Covalent binding of polychlorinated biphenyls to rat liver microsomes in vitro: nature of reactive metabolites and target macromolecules. Toxicol Appl Pharmacol 55: 490–500
Shimada T, Sawabe Y (1983) Activation of 3,4,3′,4′-tetrachlorobiphenyl to protein-bound metabolites by rat liver microsomal cytochrome P-450-containing monooxygenase system. Toxicol Appl Pharmacol 70: 486–493
Shimada T, Sawabe Y (1984) Comparative studies on distribution and covalent tissue binding of 2,4,2′, 4′- and 3,4,3′,4′-tetrachlorobiphenyl isomers in the rat. Arch Toxicol 55: 182–185
Standnicki SS, Allen JR (1979) Toxicity of 2,2′,5,5′-tetrachlorobiphenyl and its metabolites, 2,2′,5,5′-tetrachlorobiphenyl-3,4-oxide and 2,2′,5,5′-tetrachlorobiphenyl-4-ol to cultured cells in vitro. Bull Environ Contam Toxicol 23: 788–796
Steck TL, Kant JA (1974) Preparation of impermeable ghosts and inside-out vesicles from human erythrocyte membranes. Method Enzymol 31: 172–180
Taniguchi H, Imai Y, Iyanagi T, Sato R (1979) Interaction between NADPH-cytochrome P-450 reductase and cytochrome P-450 in the membrane of phosphatidylcholine vesicles. Biochim Biophys Acta 550: 341–356
Wieland E, Neuman HG (1978) Methemoglobin formation and binding to blood constituents as indicators for the formation, availability and reactivity of activated metabolites derived from trans-4-aminostilbene and related aromatic amines. Arch Toxicol 40: 17–35
Wildenauer DB, Weger N (1979) Reactions of the trifunctional nitrogen mustard tris(2-chloroethyl)amine (HN3) with human erythrocyte membranes in vitro. Biochem Pharmacol 28: 2761–2769
Wildenauer DB, Oehlmann CE (1982) Interaction of cyclophosphamide metabolites with membrane proteins: an in vitro study with rabbit liver microsomes and human red blood cells: Effect of thiols. Biochem Pharmacol 31: 3535–3541
Yasukochi Y, Masters BSS (1976) Some properties of a detergent solubilized NADPH-cytochrome c (P-450) reductase purified by biospecific affinity chromatography. J Biol Chem 251: 5337–5344
Yoshimura Y, Yoshihara S, Koga N, Kawano K, Nagata K., Wada I, Yamauchi Y (1981) Studies on the experimental PCB poisoning in rhesus and crab eating monkeys. II. Fukuoka Acta Med 72: 155–184
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Shimada, T., Sawabe, Y. & Nakano, Y. Interaction of 3,4,3′,4′-tetrachlorobiphenyl metabolites formed by cytochrome P-450 in vitro with rat erythrocytes. Arch Toxicol 58, 20–26 (1985). https://doi.org/10.1007/BF00292611
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00292611