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. 1989 Feb;171(2):1173–1177. doi: 10.1128/jb.171.2.1173-1177.1989

Purification and properties of glutathione transferase from Issatchenkia orientalis.

H Tamaki 1, H Kumagai 1, T Tochikura 1
PMCID: PMC209716  PMID: 2914866

Abstract

Glutathione transferase (GST) (EC 2.5.1.18) was purified from a cell extract of Issatchenkia orientalis, and two GST isoenzymes were isolated. They had molecular weights of 37,500 and 40,000 and were designated GST Y-1 and GST Y-2, respectively. GST Y-1 and GST Y-2 gave single bands with molecular weights of 22,000 and 23,500, respectively, on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. GST Y-1 and GST Y-2 were immunologically distinguished from each other. GST Y-1 showed specific activity 10.4-times and 6.0-times higher when 1-chloro-2,4-dinitrobenzene and o-dinitrobenzene were used as substrates, respectively, than GST Y-2. GST activity was not detected for either isoenzyme when other substrates such as bromosulfophthalein and trans-4-phenyl-3-buten-2-one were used. GST Y-1 and GST Y-2 had Km values of 0.51 and 0.75 mM for glutathione, respectively, and of 0.16 and 4.01 mM for 1-chloro-2,4-dinitrobenzene. GST Y-1 was significantly inhibited by Cibacron blue 3G-A, and GST Y-2 was significantly inhibited by bromosulfophthalein.

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Selected References

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  1. Asaoka K. Affinity purification and characterization of glutathione S-transferases from bovine liver. J Biochem. 1984 Mar;95(3):685–696. doi: 10.1093/oxfordjournals.jbchem.a134658. [DOI] [PubMed] [Google Scholar]
  2. Asaoka K., Ito H., Takahashi K. Monkey glutathione S-aryltransferases. I. Tissue distribution and purification from the liver. J Biochem. 1977 Oct;82(4):973–981. doi: 10.1093/oxfordjournals.jbchem.a131802. [DOI] [PubMed] [Google Scholar]
  3. Asaoka K., Takahashi K. A colorimetric assay of glutathione S-transferases using o-dinitrobenzene as a substrate. J Biochem. 1983 Nov;94(5):1685–1688. [PubMed] [Google Scholar]
  4. Bach M. K., Brashler J. R., Morton D. R., Steel L. K., Kaliner M. A., Hugli T. E. Formation of leukotrienes C and D and Pharmacologic modulation of their synthesis. Adv Prostaglandin Thromboxane Leukot Res. 1982;9:103–114. [PubMed] [Google Scholar]
  5. Bensadoun A., Weinstein D. Assay of proteins in the presence of interfering materials. Anal Biochem. 1976 Jan;70(1):241–250. doi: 10.1016/s0003-2697(76)80064-4. [DOI] [PubMed] [Google Scholar]
  6. Boyland E., Chasseaud L. F. The role of glutathione and glutathione S-transferases in mercapturic acid biosynthesis. Adv Enzymol Relat Areas Mol Biol. 1969;32:173–219. doi: 10.1002/9780470122778.ch5. [DOI] [PubMed] [Google Scholar]
  7. Gawai K. R., Pawar S. S. Purification and characterization of glutathione-S-transferase from liver cytosol of phenobarbital-treated rabbits. Xenobiotica. 1984 Jul;14(7):605–607. doi: 10.3109/00498258409151456. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. 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]
  10. Habig W. H., Pabst M. J., Jakoby W. B. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem. 1974 Nov 25;249(22):7130–7139. [PubMed] [Google Scholar]
  11. Jakoby W. B. The glutathione S-transferases: a group of multifunctional detoxification proteins. Adv Enzymol Relat Areas Mol Biol. 1978;46:383–414. doi: 10.1002/9780470122914.ch6. [DOI] [PubMed] [Google Scholar]
  12. Jensson H., Alin P., Mannervik B. Glutathione transferase isoenzymes from rat liver cytosol. Methods Enzymol. 1985;113:504–507. doi: 10.1016/s0076-6879(85)13066-1. [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. Lawrence R. A., Burk R. F. Glutathione peroxidase activity in selenium-deficient rat liver. Biochem Biophys Res Commun. 1976 Aug 23;71(4):952–958. doi: 10.1016/0006-291x(76)90747-6. [DOI] [PubMed] [Google Scholar]
  16. Lee C. Y., Johnson L., Cox R. H., McKinney J. D., Lee S. M. Mouse liver glutathione S-transferases. Biochemical and immunological characterization. J Biol Chem. 1981 Aug 10;256(15):8110–8116. [PubMed] [Google Scholar]
  17. 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]
  18. Meijer J., DePierre J. W., Rannug U. Measurement of drug-metabolizing systems in Salmonella typhimurium strains G46, TA15135, TA100, TA1538 and TA98. Chem Biol Interact. 1980 Sep;31(3):247–254. doi: 10.1016/0009-2797(80)90013-7. [DOI] [PubMed] [Google Scholar]
  19. Tamura H., Ui N. A new buffer system for disc electrophoresis suitable for slightly basic proteins. J Biochem. 1972 Mar;71(3):543–545. [PubMed] [Google Scholar]
  20. WAELSCH H. Certain aspects of intermediary metabolism of glutamine, asparagine, and glutathione. Adv Enzymol Relat Subj Biochem. 1952;13:237–319. doi: 10.1002/9780470122587.ch7. [DOI] [PubMed] [Google Scholar]

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