Abstract
Mononuclear lymphocytes were isolated from the blood of 12 individuals, who had been exposed to the vapour of the soil fumigant 1,3-dichloropropene (DCP). Western blot experiments were performed on the crude lymphocyte homogenates, using a monoclonal antibody against human hepatic glutathione S-transferase (GST) isoenzyme μ, to determine the presence or absence of mu-class isoenzymes μ, and/or ψ. Nine of the individuals were found to be positive for μ and/or ψ, the remaining three individuals being negative. In addition, all individuals showed a positive staining on immunoblot of a protein of somewhat lower molecular mass than the hepatic standard. This protein was bound by the S-hexylglutathione affinity column, and presumably constitutes a new mu-class isoenzyme, which is not subject to genetic polymorphism. Determination of the specific activities of individual human GST isoenzymes towards Z-(cis-) and E-(trans-)-DCP demonstrated that mu-class isoenzymes show a considerably higher specific activity with Z-DCP than alpha-class or pi-class isoenzymes. In addition, mu-class isoenzymes were found to be 2- to 3-fold more active with Z-DCP than with E-DCP. Their activity towards E-DCP was similar to the specific activity of alpha-class isoenzymes. Genetic polymorphism for mu-class isoenzymes could thus be a determinant in the extent of excretion of mercapturic acids from Z- and E-DCP. The urinary excretion of Z- and E-DCP mercapturic acids and the respiratory exposure to Z- and E-DCP were determined for nine and eight phenotyped individuals, respectively. Urinary excretion levels (corrected for the time weighted average 8-h exposure), the urinary ratio and the elimination half-lives of the mercapturic acids of Z- and E-DCP were compared with the data on the mu-phenotype. No statistically significant differences were observed between mu-class positive and mu-class negative individuals.
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References
Board PG (1981) Biochemical genetics of glutathione S-transferase in man. Am J Genet 33: 36–43
Board PG, Suzuki T, Shaw DC (1988) Human muscle glutathione S-transferase (GST-4) shows close homology to human liver GST-1. Biochim Biophys Acta 953: 214–217
Brouwer DH, Brouwer EJ, De Vreede, JAF, Van Welie RTH, Vermeulen NPE, Van Hemmen JJ (1990) Occupational exposure to 1,3-dichloropropene in the Dutch flower bulb culture. Part I. Environmental monitoring. Arch Environ Contam. Toxicol (in press)
Carmichael J, Forrester LM, Lewis AD, Hayes JD, Hayes PC, Wolf CR (1988) Glutathione S-transferase isoenzymes and glutathione peroxidase activity in normal and tumour samples from human lung. Carcinogenesis 9: 1617–1621
Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases. The first step in mercapturic acid formation. J Biol Chem 249: 7130–7139
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275
Mannervik B (1985) The isoenzymes of glutathione transferase. Adv Enzymol 57: 357–417
Mannervik B, Alin P, Guthenberg C, Jensson H, Tahir MK, Warholm M, Jornvall H (1985) Identification of three classes of cytosolic glutathione transferase common to several mammalian species: correlation between structural data and enzymatic properties. Proc Natl Acad Sci USA 82: 7202–7206
Onkenhout W, Mulder PPJ, Bogaard PJ, Buijs W, Vermeulen NPE (1986) Identification and quantitative determination of mercapturic acids formed from Z- and E-1,3-dichloropropene by the rat, using gas chromatography with three different detection techniques. Arch Toxicol 59: 235–241
Osterloh JD, Cohen BS, Popendorf W, Pond SM (1984) Urinary excretion of the N-acetyl cysteine conjugate ofcis-1,3-dichloropropene by exposed individuals. Arch Environ Health 39: 271–275
Peters WHM, Allebes WA, Jansen PLM, Poels LG, Capel PJA (1987) Characterization and tissue specificity of a monoclonal antibody against human uridine 5′-diphosphate-glucuronosyltransferase. Gastroenterology 93: 162–169
Seidegard J, DePierre JW, Pero RW (1985) Hereditary interindividual differences in the glutathione transferase activity towards trans-stilbene oxide in resting human mononuclear leukocytes are due to particular isozyme (s). Carcinogenesis 6: 1211–1216
Seidegard J, Pero RW, Miller DG, Beattie EJ (1986) A glutathione transferase in human leukocytes as a marker for the susceptibility to lung cancer. Carcinogenesis 7: 751–753
Seidegard J, Guthenberg C, Pero RW, Mannervik B (1987) The transstilbene oxide-active glutathione transferase in human mononuclear leucocytes is identical with the hepatic glutathione transferase. Biochem J 246: 783–785
Strange RC, Faulder CG, Davis BA, Hume R, Brown JAH, Cotton W, Hopkinson DA (1984) The human glutathione S-transferases: studies on the tissue distribution and genetic variation of the GST 1, GST 2 and GST 3 isozymes. Ann Hum Genet 48: 11–20
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354
Van Bladeren PJ (1988) Formation of toxic metabolites from drugs and other xenobiotics by glutathione conjugation. TIPS 9: 295–298
Van Ommen B, Bogaards JJP, Peters WHM, Blaauboer B, Van Bladeren PJ (1990) Human hepatic glutathione S-transferases: quantification, characterization of mu class genetic polymorphism and hetero/homodimers. Biochem J (in press)
Van Welie RTH, Van Duyn P, Vermeulen NPE (1989) Determination of two mercapturic acid metabolites of 1,3-dichloropropene in human urine with gas chromatography and sulfur selective detection. J Chromatogr 496: 463–471
Van Welie RTH, Van Duyn P, Brouwer DH, Van Hemmen JJ, Brouwer EJ, Vermeulen NPE (1990) Occupational exposure to dichloropropene in the Dutch flower bulb culture. Part II. Biological monitoring by measurement of urinary excretion of two mercapturic acid metabolites. Arch Environ Contam Toxicol (in press)
Vos RME, Snoek MC, Van Berkel WJH, Muller F, Van Bladeren PJ (1988) Differential induction of rat glutathione S-transferase isoenzymes by hexachlorobenzene and benzyl isothiocyanate. Comparison with induction by phenobarbital and 3-methylcholanthrene. Biochem Pharmacol 37: 1077–1082
Warholm M, Guthenberg C, Mannervik B, Von Bahr C (1981) Purification of a new glutathione S-transferase from human liver having high activity with benzo(a)pyrene-4,5-oxide. Biochem Biophys Res Commun 98: 512–519
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Vos, R.M.E., van Welie, R.T.H., Peters, W.H.M. et al. Genetic deficiency of human class mu glutathione S-transferase isoenzymes in relation to the urinary excretion of the mercapturic acids of Z- and E-1,3-dichloropropene. Arch Toxicol 65, 95–99 (1991). https://doi.org/10.1007/BF02034933
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DOI: https://doi.org/10.1007/BF02034933