Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1992 Jun 1;284(Pt 2):313–319. doi: 10.1042/bj2840313

Cloning and heterologous expression of cDNA encoding class alpha rat glutathione transferase 8-8, an enzyme with high catalytic activity towards genotoxic alpha,beta-unsaturated carbonyl compounds.

G Stenberg 1, M Ridderström 1, A Engström 1, S E Pemble 1, B Mannervik 1
PMCID: PMC1132639  PMID: 1599415

Abstract

A cDNA clone, lambda GTRA8, encoding rat glutathione transferase subunit 8 has been isolated from a lambda gt10 rat hepatoma cDNA library. The previously known amino acid sequence of the enzyme was used to design primers for a polymerase chain reaction that yielded a 0.3 kb DNA fragment from the hepatoma library. The 0.3 kb fragment was used as a probe for screening and a 0.9 kb cDNA clone containing a complete open reading frame was obtained. After DNA sequencing and subcloning into an expression vector, the enzyme was expressed in Escherichia coli and purified. Specific activities and kcat./Km values were determined for a number of substrates, including alpha,beta-unsaturated carbonyl compounds. The highest activity was obtained with 4-hydroxyalkenals and with acrolein, genotoxic products of lipid peroxidation. In addition, the rat class Alpha glutathione transferase 8-8 displays high catalytic activity in the reaction between glutathione and the diuretic drug ethacrynic acid, a compound normally considered as a substrate characteristic for class Pi glutathione transferases.

Full text

PDF
313

Selected References

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

  1. Adang A. E., Meyer D. J., Brussee J., Van der Gen A., Ketterer B., Mulder G. J. Interaction of rat glutathione S-transferases 7-7 and 8-8 with gamma-glutamyl- or glycyl-modified glutathione analogues. Biochem J. 1989 Dec 15;264(3):759–764. doi: 10.1042/bj2640759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alin P., Danielson U. H., Mannervik B. 4-Hydroxyalk-2-enals are substrates for glutathione transferase. FEBS Lett. 1985 Jan 7;179(2):267–270. doi: 10.1016/0014-5793(85)80532-9. [DOI] [PubMed] [Google Scholar]
  3. Alin P., Jensson H., Cederlund E., Jörnvall H., Mannervik B. Cytosolic glutathione transferases from rat liver. Primary structure of class alpha glutathione transferase 8-8 and characterization of low-abundance class Mu glutathione transferases. Biochem J. 1989 Jul 15;261(2):531–539. doi: 10.1042/bj2610531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benedetti A., Comporti M., Esterbauer H. Identification of 4-hydroxynonenal as a cytotoxic product originating from the peroxidation of liver microsomal lipids. Biochim Biophys Acta. 1980 Nov 7;620(2):281–296. doi: 10.1016/0005-2760(80)90209-x. [DOI] [PubMed] [Google Scholar]
  5. Benson A. M., Talalay P., Keen J. H., Jakoby W. B. Relationship between the soluble glutathione-dependent delta 5-3-ketosteroid isomerase and the glutathione S-transferases of the liver. Proc Natl Acad Sci U S A. 1977 Jan;74(1):158–162. doi: 10.1073/pnas.74.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Berhane K., Mannervik B. Inactivation of the genotoxic aldehyde acrolein by human glutathione transferases of classes alpha, mu, and pi. Mol Pharmacol. 1990 Feb;37(2):251–254. [PubMed] [Google Scholar]
  7. Board P. G., Webb G. C. Isolation of a cDNA clone and localization of human glutathione S-transferase 2 genes to chromosome band 6p12. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2377–2381. doi: 10.1073/pnas.84.8.2377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Daniel V., Sharon R., Tichauer Y., Sarid S. Mouse glutathione S-transferase Ya subunit: gene structure and sequence. DNA. 1987 Aug;6(4):317–324. doi: 10.1089/dna.1987.6.317. [DOI] [PubMed] [Google Scholar]
  9. Danielson U. H., Esterbauer H., Mannervik B. Structure-activity relationships of 4-hydroxyalkenals in the conjugation catalysed by mammalian glutathione transferases. Biochem J. 1987 Nov 1;247(3):707–713. doi: 10.1042/bj2470707. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Danielson U. H., Mannervik B. Kinetic independence of the subunits of cytosolic glutathione transferase from the rat. Biochem J. 1985 Oct 15;231(2):263–267. doi: 10.1042/bj2310263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dirr H. W., Schabort J. C. Glutathione S-transferase composition of rat erythrocytes. Biochem Int. 1987 Aug;15(2):381–384. [PubMed] [Google Scholar]
  12. Esterbauer H., Cheeseman K. H., Dianzani M. U., Poli G., Slater T. F. Separation and characterization of the aldehydic products of lipid peroxidation stimulated by ADP-Fe2+ in rat liver microsomes. Biochem J. 1982 Oct 15;208(1):129–140. doi: 10.1042/bj2080129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Flinta C., Persson B., Jörnvall H., von Heijne G. Sequence determinants of cytosolic N-terminal protein processing. Eur J Biochem. 1986 Jan 2;154(1):193–196. doi: 10.1111/j.1432-1033.1986.tb09378.x. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. 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]
  16. Hayes J. D. Purification and physical characterization of glutathione S-transferase K. Differential use of S-hexylglutathione and glutathione affinity matrices to isolate a novel glutathione S-transferase from rat liver. Biochem J. 1986 Feb 1;233(3):789–798. doi: 10.1042/bj2330789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. 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]
  20. Lathe R. Synthetic oligonucleotide probes deduced from amino acid sequence data. Theoretical and practical considerations. J Mol Biol. 1985 May 5;183(1):1–12. doi: 10.1016/0022-2836(85)90276-1. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Mankowitz L., Castro V. M., Mannervik B., Rydström J., DePierre J. W. Increase in the amount of glutathione transferase 4-4 in the rat adrenal gland after hypophysectomy and down-regulation by subsequent treatment with adrenocorticotrophic hormone. Biochem J. 1990 Jan 1;265(1):147–154. doi: 10.1042/bj2650147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Mannervik B. Glutathione peroxidase. Methods Enzymol. 1985;113:490–495. doi: 10.1016/s0076-6879(85)13063-6. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Meyer D. J., Lalor E., Coles B., Kispert A., Alin P., Mannervik B., Ketterer B. Single-step purification and h.p.l.c. analysis of glutathione transferase 8-8 in rat tissues. Biochem J. 1989 Jun 15;260(3):785–788. doi: 10.1042/bj2600785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Murugesan N., Xu C., Ehrenfeld G. M., Sugiyama H., Kilkuskie R. E., Rodriguez L. O., Chang L. H., Hecht S. M. Analysis of products formed during bleomycin-mediated DNA degradation. Biochemistry. 1985 Oct 8;24(21):5735–5744. doi: 10.1021/bi00342a008. [DOI] [PubMed] [Google Scholar]
  29. Pemble S. E., Taylor J. B., Ketterer B. Tissue distribution of rat glutathione transferase subunit 7, a hepatoma marker. Biochem J. 1986 Dec 15;240(3):885–889. doi: 10.1042/bj2400885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pickett C. B., Lu A. Y. Glutathione S-transferases: gene structure, regulation, and biological function. Annu Rev Biochem. 1989;58:743–764. doi: 10.1146/annurev.bi.58.070189.003523. [DOI] [PubMed] [Google Scholar]
  31. 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]
  32. Rhoads D. M., Zarlengo R. P., Tu C. P. The basic glutathione S-transferases from human livers are products of separate genes. Biochem Biophys Res Commun. 1987 May 29;145(1):474–481. doi: 10.1016/0006-291x(87)91345-3. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Simons P. C., Vander Jagt D. L. Purification of glutathione S-transferases from human liver by glutathione-affinity chromatography. Anal Biochem. 1977 Oct;82(2):334–341. doi: 10.1016/0003-2697(77)90169-5. [DOI] [PubMed] [Google Scholar]
  36. Stenberg G., Board P. G., Carlberg I., Mannervik B. Effects of directed mutagenesis on conserved arginine residues in a human Class Alpha glutathione transferase. Biochem J. 1991 Mar 1;274(Pt 2):549–555. doi: 10.1042/bj2740549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Telakowski-Hopkins C. A., Rodkey J. A., Bennett C. D., Lu A. Y., Pickett C. B. Rat liver glutathione S-transferases. Construction of a cDNA clone complementary to a Yc mRNA and prediction of the complete amino acid sequence of a Yc subunit. J Biol Chem. 1985 May 10;260(9):5820–5825. [PubMed] [Google Scholar]
  38. Telakowski-Hopkins C. A., Rothkopf G. S., Pickett C. B. Structural analysis of a rat liver glutathione S-transferase Ya gene. Proc Natl Acad Sci U S A. 1986 Dec;83(24):9393–9397. doi: 10.1073/pnas.83.24.9393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tu C. P., Qian B. Human liver glutathione S-transferases: complete primary sequence of an Ha subunit cDNA. Biochem Biophys Res Commun. 1986 Nov 26;141(1):229–237. doi: 10.1016/s0006-291x(86)80358-8. [DOI] [PubMed] [Google Scholar]
  40. Widersten M., Pearson W. R., Engström A., Mannervik B. Heterologous expression of the allelic variant mu-class glutathione transferases mu and psi. Biochem J. 1991 Jun 1;276(Pt 2):519–524. doi: 10.1042/bj2760519. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES