Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1988 Aug 1;253(3):759–764. doi: 10.1042/bj2530759

Isoenzymes of glutathione transferase in rat small intestine.

M K Tahir 1, N Ozer 1, B Mannervik 1
PMCID: PMC1149368  PMID: 3140787

Abstract

The major glutathione transferases in the rat small-intestine cytosol were isolated and characterized. The enzymes active with 1-chloro-2,4-dinitrobenzene as second substrate were almost quantitatively recovered after affinity chromatography on immobilized S-hexylglutathione. The different basic forms of glutathione transferase, which account for 90% of the activity, were resolved by chromatofocusing. Fractions containing enzymes with lower isoelectric points were not further resolved. The isolated fractions were characterized by their elution position in chromatofocusing, apparent subunit Mr, reactions with specific antibodies, substrate specificities and inhibition characteristics. The major basic forms identified were glutathione transferases 1-1, 4-4 and 7-7. In addition, evidence for the presence of a variant form of subunit 1, as well as trace amounts of subunits 2 and 3, was obtained. A significant amount of transferase 8-8 in the fraction of acidic enzyme forms was demonstrated by immunoblot and Ouchterlony double-diffusion analysis. In the comparison of the occurrence of the different forms of glutathione transferase in liver, lung, kidney and small intestine, it was found that the small intestine is the richest source of glutathione transferase 7-7.

Full text

PDF
759

Images in this article

761Fig. 2.
on p.761
761Fig. 3.
on p.761
762Fig. 4.
on p.762

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alin P., Jensson H., Guthenberg C., Danielson U. H., Tahir M. K., Mannervik B. Purification of major basic glutathione transferase isoenzymes from rat liver by use of affinity chromatography and fast protein liquid chromatofocusing. Anal Biochem. 1985 May 1;146(2):313–320. doi: 10.1016/0003-2697(85)90545-7. [DOI] [PubMed] [Google Scholar]
  2. Beale D., Ketterer B., Carne T., Meyer D., Taylor J. B. Evidence that the Ya and Yc subunits of glutathione transferase B (ligandin) are the products of separate genes. Eur J Biochem. 1982 Sep 1;126(3):459–463. doi: 10.1111/j.1432-1033.1982.tb06802.x. [DOI] [PubMed] [Google Scholar]
  3. Berzins K., Wahlgren M., Perlmann P. Studies on the specificity of anti-erythrocyte antibodies in the serum of patients with malaria. Clin Exp Immunol. 1983 Nov;54(2):313–318. [PMC free article] [PubMed] [Google Scholar]
  4. Chasseaud L. F. The role of glutathione and glutathione S-transferases in the metabolism of chemical carcinogens and other electrophilic agents. Adv Cancer Res. 1979;29:175–274. doi: 10.1016/s0065-230x(08)60848-9. [DOI] [PubMed] [Google Scholar]
  5. Clifton G., Kaplowitz N. Effect of dietary phenobarbital, 3,4-benzo(alpha)pyrene and 3-methylcholanthrene on hepatic, intestinal and renal glutathione s-transferase activities in the rat. Biochem Pharmacol. 1978;27(8):1284–1287. doi: 10.1016/0006-2952(78)90465-3. [DOI] [PubMed] [Google Scholar]
  6. Guthenberg C., Alin P., Mannervik B. Glutathione transferase from rat testis. Methods Enzymol. 1985;113:507–510. doi: 10.1016/s0076-6879(85)13067-3. [DOI] [PubMed] [Google Scholar]
  7. Guthenberg C., Jensson H., Nyström L., Osterlund E., Tahir M. K., Mannervik B. Isoenzymes of glutathione transferase in rat kidney cytosol. Biochem J. 1985 Sep 15;230(3):609–615. doi: 10.1042/bj2300609. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Guthenberg C., Mannervik B. Glutathione S-transferase (transferase pi) from human placenta is identical or closely related to glutathione S-transferase (transferase rho) from erythrocytes. Biochim Biophys Acta. 1981 Oct 13;661(2):255–260. doi: 10.1016/0005-2744(81)90012-7. [DOI] [PubMed] [Google Scholar]
  9. Hayes J. D., Mantle T. J. Use of immuno-blot techniques to discriminate between the glutathione S-transferase Yf, Yk, Ya, Yn/Yb and Yc subunits and to study their distribution in extrahepatic tissues. Evidence for three immunochemically distinct groups of transferase in the rat. Biochem J. 1986 Feb 1;233(3):779–788. doi: 10.1042/bj2330779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jakoby W. B., Ketterer B., Mannervik B. Glutathione transferases: nomenclature. Biochem Pharmacol. 1984 Aug 15;33(16):2539–2540. doi: 10.1016/0006-2952(84)90621-x. [DOI] [PubMed] [Google Scholar]
  11. Jensson H., Eriksson L. C., Mannervik B. Selective expression of glutathione transferase isoenzymes in chemically induced preneoplastic rat hepatocyte nodules. FEBS Lett. 1985 Jul 22;187(1):115–120. doi: 10.1016/0014-5793(85)81225-4. [DOI] [PubMed] [Google Scholar]
  12. Jensson H., Guthenberg C., Alin P., Mannervik B. Rat glutathione transferase 8-8, an enzyme efficiently detoxifying 4-hydroxyalk-2-enals. FEBS Lett. 1986 Jul 28;203(2):207–209. doi: 10.1016/0014-5793(86)80743-8. [DOI] [PubMed] [Google Scholar]
  13. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  14. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  15. Lai H. C., Li N., Weiss M. J., Reddy C. C., Tu C. P. The nucleotide sequence of a rat liver glutathione S-transferase subunit cDNA clone. J Biol Chem. 1984 May 10;259(9):5536–5542. [PubMed] [Google Scholar]
  16. Mannervik B., Alin P., Guthenberg C., Jensson H., Tahir M. K., Warholm M., Jörnvall H. Identification of three classes of cytosolic glutathione transferase common to several mammalian species: correlation between structural data and enzymatic properties. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7202–7206. doi: 10.1073/pnas.82.21.7202. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Mannervik B., Guthenberg C. Glutathione transferase (human placenta). Methods Enzymol. 1981;77:231–235. doi: 10.1016/s0076-6879(81)77030-7. [DOI] [PubMed] [Google Scholar]
  18. Mannervik B., Jensson H. Binary combinations of four protein subunits with different catalytic specificities explain the relationship between six basic glutathione S-transferases in rat liver cytosol. J Biol Chem. 1982 Sep 10;257(17):9909–9912. [PubMed] [Google Scholar]
  19. Mannervik B. The isoenzymes of glutathione transferase. Adv Enzymol Relat Areas Mol Biol. 1985;57:357–417. doi: 10.1002/9780470123034.ch5. [DOI] [PubMed] [Google Scholar]
  20. Meyer D. J., Beale D., Tan K. H., Coles B., Ketterer B. Glutathione transferases in primary rat hepatomas: the isolation of a form with GSH peroxidase activity. FEBS Lett. 1985 May 6;184(1):139–143. doi: 10.1016/0014-5793(85)80670-0. [DOI] [PubMed] [Google Scholar]
  21. Meyer D. J., Christodoulides L. G., Hong Tan K., Ketterer B. Isolation, properties and tissue distribution of rat glutathione transferase E. FEBS Lett. 1984 Aug 6;173(2):327–330. doi: 10.1016/0014-5793(84)80799-1. [DOI] [PubMed] [Google Scholar]
  22. Ogasawara T., Hoensch H., Ohnhaus E. E. Distribution of glutathione and its related enzymes in small intestinal mucosa of rats. Arch Toxicol Suppl. 1985;8:110–113. doi: 10.1007/978-3-642-69928-3_13. [DOI] [PubMed] [Google Scholar]
  23. Ostlund Farrants A. K., Meyer D. J., Coles B., Southan C., Aitken A., Johnson P. J., Ketterer B. The separation of glutathione transferase subunits by using reverse-phase high-pressure liquid chromatography. Biochem J. 1987 Jul 15;245(2):423–428. doi: 10.1042/bj2450423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Pickett C. B., Telakowski-Hopkins C. A., Ding G. J., Argenbright L., Lu A. Y. Rat liver glutathione S-transferases. Complete nucleotide sequence of a glutathione S-transferase mRNA and the regulation of the Ya, Yb, and Yc mRNAs by 3-methylcholanthrene and phenobarbital. J Biol Chem. 1984 Apr 25;259(8):5182–5188. [PubMed] [Google Scholar]
  25. Pinkus L. M., Ketley J. N., Jakoby W. B. The glutathione S-transferases as a possible detoxification system of rat intestinal epithelium. Biochem Pharmacol. 1977 Dec 15;26(24):2359–2363. doi: 10.1016/0006-2952(77)90441-5. [DOI] [PubMed] [Google Scholar]
  26. Robertson I. G., Jensson H., Guthenberg C., Tahir M. K., Jernström B., Mannervik B. Differences in the occurrence of glutathione transferase isoenzymes in rat lung and liver. Biochem Biophys Res Commun. 1985 Feb 28;127(1):80–86. doi: 10.1016/s0006-291x(85)80128-5. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Sato K., Kitahara A., Satoh K., Ishikawa T., Tatematsu M., Ito N. The placental form of glutathione S-transferase as a new marker protein for preneoplasia in rat chemical hepatocarcinogenesis. Gan. 1984 Mar;75(3):199–202. [PubMed] [Google Scholar]
  29. Satoh K., Kitahara A., Sato K. Identification of heterogeneous and microheterogeneous subunits of glutathione S-transferase in rat liver cytosol. Arch Biochem Biophys. 1985 Oct;242(1):104–111. doi: 10.1016/0003-9861(85)90484-9. [DOI] [PubMed] [Google Scholar]
  30. Satoh K., Kitahara A., Soma Y., Inaba Y., Hatayama I., Sato K. Purification, induction, and distribution of placental glutathione transferase: a new marker enzyme for preneoplastic cells in the rat chemical hepatocarcinogenesis. Proc Natl Acad Sci U S A. 1985 Jun;82(12):3964–3968. doi: 10.1073/pnas.82.12.3964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sheehan D., Mantle T. J. Evidence for two forms of ligandin (YaYa dimers of glutathione S-transferase) in rat liver and kidney. Biochem J. 1984 Mar 15;218(3):893–897. doi: 10.1042/bj2180893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wang I. Y., Tung E., Wang A. C., Argenbright L., Wang R., Pickett C. B., Lu A. Y. Multiple Ya subunits of glutathione S-transferase detected by monoclonal antibodies. Arch Biochem Biophys. 1986 Mar;245(2):543–547. doi: 10.1016/0003-9861(86)90249-3. [DOI] [PubMed] [Google Scholar]
  33. Warholm M., Guthenberg C., Mannervik B. Molecular and catalytic properties of glutathione transferase mu from human liver: an enzyme efficiently conjugating epoxides. Biochemistry. 1983 Jul 19;22(15):3610–3617. doi: 10.1021/bi00284a011. [DOI] [PubMed] [Google Scholar]
  34. Warholm M., Jensson H., Tahir M. K., Mannervik B. Purification and characterization of three distinct glutathione transferases from mouse liver. Biochemistry. 1986 Jul 15;25(14):4119–4125. doi: 10.1021/bi00362a020. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES