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. 1991 May 1;275(Pt 3):703–709. doi: 10.1042/bj2750703

The initial-rate kinetics of mouse glutathione S-transferase YfYf. Evidence for an allosteric site for ethacrynic acid.

M F Phillips 1, T J Mantle 1
PMCID: PMC1150111  PMID: 2039447

Abstract

Mouse glutathione S-transferase GST YfYf (an orthologue of GST P or 7-7 in the rat and of GST pi in the human) was found to have a subunit Mr of 24,500 and cross-reacted with anti-(rat GST YfYf). N-Terminal analysis showed a close similarity to the rat, human and bovine orthologues. On isoelectric focusing the native enzyme had a pI of 8.3 and a pI of 7.3 in the presence of urea. Initial-rate studies with 1-chloro-2,4-dinitrobenzene (CDNB) and GSH as substrates and inhibition studies with the product of the enzyme-catalysed conjugation of CDNB and GSH, S-(2,4-dinitrophenyl)glutathione, indicated a rapid-equilibrium random mechanism for the enzyme. The diuretic drug ethacrynic acid was found to be simultaneously a competitive inhibitor and an uncompetitive activator of the enzyme (with CDNB as the substrate whose concentration was varied). By using a computer simulation program (EKPLOT) a model was developed that would explain the experimental data. It is proposed that ethacrynic acid can compete with CDNB at the active site but simultaneously bind to an allosteric site on the enzyme, causing an elevation in the Vmax. for the conjugation of CDNB and GSH. The implications of such an activation mechanism for an enzyme potentially conjugating a range of xenobiotic compounds are discussed.

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Selected References

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  1. Alin P., Mannervik B., Jörnvall H. Cytosolic rat liver glutathione transferase 4-4. Primary structure of the protein reveals extensive differences between homologous glutathione transferases of classes alpha and mu. Eur J Biochem. 1986 Apr 15;156(2):343–350. doi: 10.1111/j.1432-1033.1986.tb09588.x. [DOI] [PubMed] [Google Scholar]
  2. Alin P., Mannervik B., Jörnvall H. Structural evidence for three different types of glutathione transferase in human tissues. FEBS Lett. 1985 Mar 25;182(2):319–322. doi: 10.1016/0014-5793(85)80324-0. [DOI] [PubMed] [Google Scholar]
  3. Caccuri A. M., Aceto A., Piemonte F., Di Ilio C., Rosato N., Federici G. Interaction of hemin with placental glutathione transferase. Eur J Biochem. 1990 May 20;189(3):493–497. doi: 10.1111/j.1432-1033.1990.tb15514.x. [DOI] [PubMed] [Google Scholar]
  4. Clark A. G., Carrol N. Suppression of high-affinity ligand binding to the major glutathione S-transferase from Galleria mellonella by physiological concentrations of glutathione. Biochem J. 1986 Jan 15;233(2):325–331. doi: 10.1042/bj2330325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clark A. G., Dick G. L., Smith J. N. Kinetic studies on a glutathione S-transferase from the larvae of Costelytra zealandica. Biochem J. 1984 Jan 1;217(1):51–58. doi: 10.1042/bj2170051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Corrigall A. V., Bhargava M. M., Ivanetich K. M., Ehlers M. R., Kirsch R. E. Site-directed inactivation of human lung acidic glutathione S-transferase by 1-chloro-2,4-dinitrobenzene in the absence of glutathione. Biochim Biophys Acta. 1989 Jun 27;991(3):399–404. doi: 10.1016/0304-4165(89)90064-0. [DOI] [PubMed] [Google Scholar]
  7. Danielson U. H., Mannervik B. Paradoxical inhibition of rat glutathione transferase 4-4 by indomethacin explained by substrate-inhibitor-enzyme complexes in a random-order sequential mechanism. Biochem J. 1988 Mar 15;250(3):705–711. doi: 10.1042/bj2500705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dao D. D., Partridge C. A., Kurosky A., Awasthi Y. C. Human glutathione S-transferases. Characterization of the anionic forms from lung and placenta. Biochem J. 1984 Jul 1;221(1):33–41. doi: 10.1042/bj2210033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. FRIEDEN C. Glutamic dehydrogenase. II. The effect of various nucleotides on the association-dissociation and kinetic properties. J Biol Chem. 1959 Apr;234(4):815–820. [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. Hatayama I., Satoh K., Sato K. Developmental and hormonal regulation of the major form of hepatic glutathione S-transferase in male mice. Biochem Biophys Res Commun. 1986 Oct 30;140(2):581–588. doi: 10.1016/0006-291x(86)90771-0. [DOI] [PubMed] [Google Scholar]
  12. Hayes J. D., Coulthwaite R. E., Stockman P. K., Hussey A. J., Mantle T. J., Wolf C. R. Glutathione S-transferase subunits in the mouse and their catalytic activities towards reactive electrophiles. Arch Toxicol Suppl. 1987;10:136–146. doi: 10.1007/978-3-642-71617-1_11. [DOI] [PubMed] [Google Scholar]
  13. Hayes J. D., Mantle T. J. Inhibition of hepatic and extrahepatic glutathione S-transferases by primary and secondary bile acids. Biochem J. 1986 Jan 15;233(2):407–415. doi: 10.1042/bj2330407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Ivanetich K. M., Goold R. D. A rapid equilibrium random sequential bi-bi mechanism for human placental glutathione S-transferase. Biochim Biophys Acta. 1989 Sep 14;998(1):7–13. doi: 10.1016/0167-4838(89)90111-8. [DOI] [PubMed] [Google Scholar]
  16. Iwatsubo M., Pantaloni D. Régulation de l'activité de la glutamate déshydrogènase par les effecteurs GTP et ADP: étude par "stopped flow". Bull Soc Chim Biol (Paris) 1967 Dec 18;49(11):1563–1572. [PubMed] [Google Scholar]
  17. Jagt D. L., Wilson S. P., Dean V. L., Simons P. C. Bilirubin binding to rat liver ligandins (glutathione S-transferases A and B). Relationship between bilirubin binding and transferase activity. J Biol Chem. 1982 Feb 25;257(4):1997–2001. [PubMed] [Google Scholar]
  18. Jakobson I., Askelöf P., Warholm M., Mannervik B. A steady-state-kinetic random mechanism for glutathione S-transferase A from rat liver. A model involving kinetically significant enzyme-product complexes in the forward reaction. Eur J Biochem. 1977 Jul 15;77(2):253–262. doi: 10.1111/j.1432-1033.1977.tb11664.x. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. Kano T., Sakai M., Muramatsu M. Structure and expression of a human class pi glutathione S-transferase messenger RNA. Cancer Res. 1987 Nov 1;47(21):5626–5630. [PubMed] [Google Scholar]
  22. Kitahara A., Satoh K., Nishimura K., Ishikawa T., Ruike K., Sato K., Tsuda H., Ito N. Changes in molecular forms of rat hepatic glutathione S-transferase during chemical hepatocarcinogenesis. Cancer Res. 1984 Jun;44(6):2698–2703. [PubMed] [Google Scholar]
  23. 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]
  24. Lai H. C., Qian B., Grove G., Tu C. P. Gene expression of rat glutathione S-transferases. Evidence for gene conversion in the evolution of the Yb multigene family. J Biol Chem. 1988 Aug 15;263(23):11389–11395. [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Markau K., Schneider J., Sund H. Kinetic studies on the mechanism of the action of ADP on the glutamate dehydrogenase reaction. FEBS Lett. 1972 Jul 15;24(1):32–36. doi: 10.1016/0014-5793(72)80819-6. [DOI] [PubMed] [Google Scholar]
  28. Markwell M. A., Haas S. M., Bieber L. L., Tolbert N. E. A modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples. Anal Biochem. 1978 Jun 15;87(1):206–210. doi: 10.1016/0003-2697(78)90586-9. [DOI] [PubMed] [Google Scholar]
  29. McLellan L. I., Hayes J. D. Sex-specific constitutive expression of the pre-neoplastic marker glutathione S-transferase, YfYf, in mouse liver. Biochem J. 1987 Jul 15;245(2):399–406. doi: 10.1042/bj2450399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pabst M. J., Habig W. H., Jakoby W. B. Glutathione S-transferase A. A novel kinetic mechanism in which the major reaction pathway depends on substrate concentration. J Biol Chem. 1974 Nov 25;249(22):7140–7147. [PubMed] [Google Scholar]
  31. Principato G. B., Danielson U. H., Mannervik B. Relaxed thiol substrate specificity of glutathione transferase effected by a non-substrate glutathione derivative. FEBS Lett. 1988 Apr 11;231(1):155–158. doi: 10.1016/0014-5793(88)80722-1. [DOI] [PubMed] [Google Scholar]
  32. Rothkopf G. S., Telakowski-Hopkins C. A., Stotish R. L., Pickett C. B. Multiplicity of glutathione S-transferase genes in the rat and association with a type 2 Alu repetitive element. Biochemistry. 1986 Mar 11;25(5):993–1002. doi: 10.1021/bi00353a007. [DOI] [PubMed] [Google Scholar]
  33. Schramm V. L., McCluskey R., Emig F. A., Litwack G. Kinetic studies and active site-binding properties of glutathione S-transferase using spin-labeled glutathione, a product analogue. J Biol Chem. 1984 Jan 25;259(2):714–722. [PubMed] [Google Scholar]
  34. Schäffer J., Gallay O., Ladenstein R. Glutathione transferase from bovine placenta. Preparation, biochemical characterization, crystallization, and preliminary crystallographic analysis of a neutral class PI enzyme. J Biol Chem. 1988 Nov 25;263(33):17405–17411. [PubMed] [Google Scholar]
  35. Strange R. C., Cramb R., Hayes J. D., Percy-Robb I. W. Partial purification of two lithocholic acid-binding proteins from rat liver 100 000g supernatants. Biochem J. 1977 Sep 1;165(3):425–429. doi: 10.1042/bj1650425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Suguoka Y., Kano T., Okuda A., Sakai M., Kitagawa T., Muramatsu M. Cloning and the nucleotide sequence of rat glutathione S-transferase P cDNA. Nucleic Acids Res. 1985 Sep 11;13(17):6049–6057. doi: 10.1093/nar/13.17.6049. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Vince R., Daluge S., Wadd W. B. Studies on the inhibition of glyoxalase I by S-substituted glutathiones. J Med Chem. 1971 May;14(5):402–404. doi: 10.1021/jm00287a006. [DOI] [PubMed] [Google Scholar]

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