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. 2000 Aug 1;349(Pt 3):693–696. doi: 10.1042/bj3490693

Studies on the differential inhibition of glutathione conjugate formation of (+)-anti-benzo[a]pyrene 7,8-dihydrodiol 9,10-epoxide and 1-chloro-2,4-dinitrobenzene in V79 Chinese hamster cells.

K Sundberg 1, B Jernström 1, S Swedmark 1
PMCID: PMC1221194  PMID: 10903128

Abstract

V79 Chinese hamster cells have previously been shown to lack the capacity to detoxify the mutagenic and carcinogenic compound (+)-anti-benzo[a]pyrene 7,8-dihydrodiol 9,10-epoxide [(+)-anti-BPDE] by Pi class glutathione transferase (GSTPi)-catalysed conjugation with GSH, although these cells contain such an enzyme [Romert, Dock, Jenssen and Jernström (1989) Carcinogenesis 10, 1701-1707; Swedmark, Romert, Morgenstern and Jenssen (1992) Carcinogenesis 13, 1719-1723; Swedmark and Jenssen (1994) Gene 139, 251-256]. Previous findings also indicate that these results do not depend on an inactive GSTPi enzyme, since V79 cells conjugate 1-chloro-2, 4-dinitrobenzene (CDNB) with GSH, but more likely on (a) factor(s) that inhibit(s) V79 GSTPi selectively [Swedmark, Jernström and Jenssen (1996) Biochem. J. 318, 533-538]. The present study demonstrates that both human and V79 recombinant GSTPi enzymes are inhibited with respect to conjugating (+)-anti-BPDE, but not CDNB, after pre-incubation with V79-cell extract, but not with MCF-7-cell extract. In addition, it was found that the inhibition is dependent on the amount of cell extract present and that the factor(s) is heat-resistant and has a molecular mass of less than 10 kDa, suggesting that the factor(s) is (are) non-proteinaceous in nature.

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Selected References

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  1. Board P. G., Webb G. C., Coggan M. Isolation of a cDNA clone and localization of the human glutathione S-transferase 3 genes to chromosome bands 11q13 and 12q13-14. Ann Hum Genet. 1989 Jul;53(Pt 3):205–213. doi: 10.1111/j.1469-1809.1989.tb01786.x. [DOI] [PubMed] [Google Scholar]
  2. Cameron A. D., Sinning I., L'Hermite G., Olin B., Board P. G., Mannervik B., Jones T. A. Structural analysis of human alpha-class glutathione transferase A1-1 in the apo-form and in complexes with ethacrynic acid and its glutathione conjugate. Structure. 1995 Jul 15;3(7):717–727. doi: 10.1016/s0969-2126(01)00206-4. [DOI] [PubMed] [Google Scholar]
  3. Frantz C. N., Malling H. V. Bromodeoxyuridine resistance induced in mouse lymphoma cells by microsomal activation of dimethylnitrosamine. J Toxicol Environ Health. 1976 Sep;2(1):179–187. doi: 10.1080/15287397609529425. [DOI] [PubMed] [Google Scholar]
  4. Habig W. H., Jakoby W. B. Assays for differentiation of glutathione S-transferases. Methods Enzymol. 1981;77:398–405. doi: 10.1016/s0076-6879(81)77053-8. [DOI] [PubMed] [Google Scholar]
  5. Hayes J. D., Pulford D. J. The glutathione S-transferase supergene family: regulation of GST and the contribution of the isoenzymes to cancer chemoprotection and drug resistance. Crit Rev Biochem Mol Biol. 1995;30(6):445–600. doi: 10.3109/10409239509083491. [DOI] [PubMed] [Google Scholar]
  6. Jernstrom B., Funk M., Frank H., Mannervik B., Seidel A. Glutathione S-transferase A1-1-catalysed conjugation of bay and fjord region diol epoxides or polycyclic aromatic hydrocarbons with glutathione. Carcinogenesis. 1996 Jul;17(7):1491–1498. doi: 10.1093/carcin/17.7.1491. [DOI] [PubMed] [Google Scholar]
  7. Ketterer B., Christodoulides L. G. Enzymology of cytosolic glutathione S-transferases. Adv Pharmacol. 1994;27:37–69. doi: 10.1016/s1054-3589(08)61029-7. [DOI] [PubMed] [Google Scholar]
  8. Kolm R. H., Stenberg G., Widersten M., Mannervik B. High-level bacterial expression of human glutathione transferase P1-1 encoded by semisynthetic DNA. Protein Expr Purif. 1995 Jun;6(3):265–271. doi: 10.1006/prep.1995.1034. [DOI] [PubMed] [Google Scholar]
  9. Krahn D. F., Heidelberger C. Liver homogenate-mediated mutagenesis in chinese hamster V79 cells by polycyclic aromatic hydrocarbons and aflatoxins. Mutat Res. 1977 Feb 1;46(1):27–44. doi: 10.1016/0165-1161(77)90108-x. [DOI] [PubMed] [Google Scholar]
  10. Kuroki T., Drevon C., Montesano R. Microsome-mediated mutagenesis in V79 Chinese hamster cells by various nitrosamines. Cancer Res. 1977 Apr;37(4):1044–1050. [PubMed] [Google Scholar]
  11. Mannervik B., Danielson U. H. Glutathione transferases--structure and catalytic activity. CRC Crit Rev Biochem. 1988;23(3):283–337. doi: 10.3109/10409238809088226. [DOI] [PubMed] [Google Scholar]
  12. Mitra A., Govindwar S., Joseph P., Kulkarni A. Inhibition of human term placental and fetal liver glutathione-S-transferases by fatty acids and fatty acid esters. Toxicol Lett. 1992 May;60(3):281–288. doi: 10.1016/0378-4274(92)90286-s. [DOI] [PubMed] [Google Scholar]
  13. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  14. Robertson I. G., Guthenberg C., Mannervik B., Jernström B. Differences in stereoselectivity and catalytic efficiency of three human glutathione transferases in the conjugation of glutathione with 7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-oxy-7,8,9,10-tetrahydrobenzo(a)pyrene. Cancer Res. 1986 May;46(5):2220–2224. [PubMed] [Google Scholar]
  15. Robertson I. G., Jensson H., Mannervik B., Jernström B. Glutathione transferases in rat lung: the presence of transferase 7-7, highly efficient in the conjugation of glutathione with the carcinogenic (+)-7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene. Carcinogenesis. 1986 Feb;7(2):295–299. doi: 10.1093/carcin/7.2.295. [DOI] [PubMed] [Google Scholar]
  16. Romert L., Dock L., Jenssen D., Jernström B. Effects of glutathione transferase activity on benzo[a]pyrene 7,8-dihydrodiol metabolism and mutagenesis studied in a mammalian cell co-cultivation assay. Carcinogenesis. 1989 Sep;10(9):1701–1707. doi: 10.1093/carcin/10.9.1701. [DOI] [PubMed] [Google Scholar]
  17. Stenberg G., Björnestedt R., Mannervik B. Heterologous expression of recombinant human glutathione transferase A1-1 from a hepatoma cell line. Protein Expr Purif. 1992 Feb;3(1):80–84. doi: 10.1016/1046-5928(92)90060-a. [DOI] [PubMed] [Google Scholar]
  18. Sundberg K., Widersten M., Seidel A., Mannervik B., Jernström B. Glutathione conjugation of bay- and fjord-region diol epoxides of polycyclic aromatic hydrocarbons by glutathione transferases M1-1 and P1-1. Chem Res Toxicol. 1997 Nov;10(11):1221–1227. doi: 10.1021/tx970099w. [DOI] [PubMed] [Google Scholar]
  19. Swedmark S., Jenssen D. Sequence of the mRNA for a glutathione transferase Pi with a different substrate specificity in V79 Chinese hamster lung cells. Gene. 1994 Feb 25;139(2):251–256. doi: 10.1016/0378-1119(94)90765-x. [DOI] [PubMed] [Google Scholar]
  20. Swedmark S., Jernström B., Jenssen D. Comparison of the mRNA sequences for Pi class glutathione transferases in different hamster species and the corresponding enzyme activities with anti-benzo[a]pyrene-7,8-dihydrodiol 9,10-epoxide. Biochem J. 1996 Sep 1;318(Pt 2):533–538. doi: 10.1042/bj3180533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Swedmark S., Romert L., Morgenstern R., Jenssen D. Studies on glutathione transferases belonging to class pi in cell lines with different capacities for conjugating (+)-7 beta, 8 alpha-dihydroxy-9 alpha, 10 alpha-oxy-7,8,9,10-tetrahydrobenzo[a]pyrene. Carcinogenesis. 1992 Oct;13(10):1719–1723. doi: 10.1093/carcin/13.10.1719. [DOI] [PubMed] [Google Scholar]

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