title: The microsomal metabolism of hexachlorobenzene. Origin of the covalent binding to protein.
abstract: The microsomal metabolism of hexachlorobenzene is studied, with special attention to the covalent binding to protein. The metabolites formed are pentachlorophenol and tetrachlorohydroquinone. In addition, a considerable amount of covalent binding to protein is detected (250 pmoles pentachlorophenol, 17 pmoles tetrachlorohydroquinone and 11 pmoles covalent binding in an incubation containing 50 mumoles of hexachlorobenzene). In order to establish the potential role of reductive dechlorination in the covalent binding, the anaerobic metabolism of hexachlorobenzene was investigated. At low oxygen concentrations no pentachlorobenzene was detected, and only very small amounts of pentachlorophenol as well as covalent binding, indicating a relationship between covalent binding and the microsomal oxidation of hexachlorobenzene. Incubations with 14C-pentachlorophenol at low concentrations showed that a conversion-dependent covalent binding occurs to the extent of 75 pmole binding per nmole pentachlorophenol. This is almost enough to account for the amount of label bound to protein observed in hexachlorobenzene incubations. This indicates that less than 10% of the covalent binding occurs during conversion of hexachlorobenzene to pentachlorophenol, and the remainder is produced during conversion of hexachlorobenzene to pentachlorophenol, and the remainder is produced during conversion of pentachlorophenol. The major product of microsomal oxidation of pentachlorophenol is tetrachlorohydroquinone, which is in redox-equilibrium with the corresponding semiquinone and quinone (chloranil). The covalent binding is inhibited by addition of ascorbic acid or glutathione to the hexachlorobenzene incubations. Ascorbic acid decreases the covalent binding with a simultaneous increase in formation of tetrachlorohydroquinone, probably due to a shift in the redox-equilibrium to the reduced side. Glutathione does not act as a reducing agent, since the inhibition of covalent binding is not accompanied by an increase in tetrachlorohydroquinone formation. Instead, glutathione reacts with chloranil, producing at least three stable products, probably in a Michael-type reaction. These results strongly indicate the involvement of chloranil or the semiquinone radical in the covalent binding during microsomal hexachlorobenzene metabolism.
mesh_terms: D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D002703:Chloranil / Q000494:pharmacology; D002722:Chlorobenzenes / Q000378:metabolism*; D005978:Glutathione / Q000494:pharmacology; D006581:Hexachlorobenzene / Q000378:metabolism*; D006873:Hydroquinones / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D008861:Microsomes / Q000378:metabolism*; D010416:Pentachlorophenol / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains
chemical_list: D002722:Chlorobenzenes; D006873:Hydroquinones; D002703:Chloranil; D006581:Hexachlorobenzene; C044578:2,3,5,6-tetrachlorohydroquinone; D010416:Pentachlorophenol; D005978:Glutathione; D001205:Ascorbic Acid