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. 1986 Aug 1;237(3):731–740. doi: 10.1042/bj2370731

Anomalous electrophoretic behaviour of the glutathione S-transferase Ya and Yk subunits isolated from man and rodents. A potential pitfall for nomenclature.

J D Hayes, T J Mantle
PMCID: PMC1147051  PMID: 3800913

Abstract

GSH S-transferases are dimeric enzymes. The subunits in the rat are resolved into six types, designated Yf, Yk, Ya, Yn, Yb and Yc, by discontinuous SDS/polyacrylamide-gel electrophoresis [Hayes (1986) Biochem. J. 233, 789-798]. The relative electrophoretic mobility of the Ya and Yk subunits is dependent on the amount of cross-linker (NN'-methylenebisacrylamide) in the resolving gel. At low degrees of cross-linking, CBis 0.6% (w/w), the Yk and Ya subunits possess a faster anodal mobility than do the Yf, Yn, Yb and Yc subunits (i.e. order of mobility Yk greater than Ya greater than Yf greater than Yn greater than Yb greater than Yc), whereas at higher degrees of cross-linking, CBis 5.0% (w/w), Yf subunits possess the fastest mobility (i.e. order of mobility Yf greater than Yk greater than or equal to Yn greater than Yb greater than or equal to Ya greater than Yc). Resolving gels that contain low concentrations of cross-linker [CBis 0.6% (w/w)] allow the resolution of a hitherto unrecognized polypeptide that is isolated by S-hexyl-GSH-Sepharose affinity chromatography. This new polypeptide, which we have designated Yb, is normally obscured by the main Yb band in resolving gels that comprise concentrations of cross-linker of at least CBis 1.6% (w/w). The Ya- and Yb-type subunits in guinea pig, mouse, hamster and man were identified by immuno-blotting and their apparent Mr values in different electrophoresis systems were determined. The Ya subunits in all species studied possess a variable cross-linker-dependent mobility during electrophoresis. Since the transferase subunits are currently classified according to their mobilities during SDS/polyacrylamide-gel electrophoresis, it is apparent that the variable electrophoretic behaviour of the Ya and Yk subunits may lead to the mis-identification of enzymes.

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Selected References

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

  1. Anderson C. W., Baum P. R., Gesteland R. F. Processing of adenovirus 2-induced proteins. J Virol. 1973 Aug;12(2):241–252. doi: 10.1128/jvi.12.2.241-252.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bass N. M., Kirsch R. E., Tuff S. A., Marks I., Saunders S. J. Ligandin heterogeneity : evidence that the two non-identical subunits are the monomers of two distinct proteins. Biochim Biophys Acta. 1977 May 27;492(1):163–175. doi: 10.1016/0005-2795(77)90223-9. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Beale D., Meyer D. J., Taylor J. B., Ketterer B. Evidence that the Yb subunits of hepatic glutathione transferases represent two different but related families of polypeptides. Eur J Biochem. 1983 Dec 1;137(1-2):125–129. doi: 10.1111/j.1432-1033.1983.tb07805.x. [DOI] [PubMed] [Google Scholar]
  5. Boyer T. D., Kenney W. C., Zakim D. Structural, functional and hybridization studies of the glutathione S-transferases of rat liver. Biochem Pharmacol. 1983 Jun 15;32(12):1843–1850. doi: 10.1016/0006-2952(83)90048-5. [DOI] [PubMed] [Google Scholar]
  6. Chang C. H., Chen J. G., Govindjee R., Ebrey T. Cation binding by bacteriorhodopsin. Proc Natl Acad Sci U S A. 1985 Jan;82(2):396–400. doi: 10.1073/pnas.82.2.396. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  9. Ding G. J., Lu A. Y., Pickett C. B. Rat liver glutathione S-transferases. Nucleotide sequence analysis of a Yb1 cDNA clone and prediction of the complete amino acid sequence of the Yb1 subunit. J Biol Chem. 1985 Oct 25;260(24):13268–13271. [PubMed] [Google Scholar]
  10. Eriksson L. C., Sharma R. N., Roomi M. W., Ho R. K., Farber E., Murray R. K. A characteristic electrophoretic pattern of cytosolic polypeptides from hepatocyte nodules generated during liver carcinogenesis in several models. Biochem Biophys Res Commun. 1983 Dec 28;117(3):740–745. doi: 10.1016/0006-291x(83)91659-5. [DOI] [PubMed] [Google Scholar]
  11. Farber E. Pre-cancerous steps in carcinogenesis. Their physiological adaptive nature. Biochim Biophys Acta. 1984;738(4):171–180. doi: 10.1016/0304-419x(83)90002-1. [DOI] [PubMed] [Google Scholar]
  12. Frey A. B., Friedberg T., Oesch F., Kreibich G. Studies on the subunit composition of rat liver glutathione S-transferases. J Biol Chem. 1983 Sep 25;258(18):11321–11325. [PubMed] [Google Scholar]
  13. Hayes J. D., Chalmers J. Bile acid inhibition of basic and neutral glutathione S-transferases in rat liver. Biochem J. 1983 Dec 1;215(3):581–588. doi: 10.1042/bj2150581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hayes J. D., Clarkson G. H. Purification and characterization of three forms of glutathione S-transferase A. A comparative study of the major YaYa-, YbYb- and YcYc-containing glutathione S-transferases. Biochem J. 1982 Dec 1;207(3):459–470. doi: 10.1042/bj2070459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hayes J. D., Gilligan D., Chapman B. J., Beckett G. J. Purification of human hepatic glutathione S-transferases and the development of a radioimmunoassay for their measurement in plasma. Clin Chim Acta. 1983 Oct 31;134(1-2):107–121. doi: 10.1016/0009-8981(83)90189-4. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Hayes J. D. Purification and characterization of glutathione S-transferases P, S and N. Isolation from rat liver of Yb1 Yn protein, the existence of which was predicted by subunit hybridization in vitro. Biochem J. 1984 Dec 15;224(3):839–852. doi: 10.1042/bj2240839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Hayes J. D., Strange R. C., Percy-Robb I. W. A study of the structures of the YaYa and YaYc glutathione S-transferases from rat liver cytosol. Evidence that the Ya monomer is responsible for lithocholate-binding activity. Biochem J. 1981 Aug 1;197(2):491–502. doi: 10.1042/bj1970491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Irwin C., O'Brien J. K., Chu P., Townsend-Parchman J. K., O'Hara P., Hunter F. E., Jr Glutathione peroxidase II of guinea pig liver cytosol: relationship to glutathione S-transferases. Arch Biochem Biophys. 1980 Nov;205(1):122–131. doi: 10.1016/0003-9861(80)90090-9. [DOI] [PubMed] [Google Scholar]
  21. 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]
  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. 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]
  25. 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]
  26. 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]
  27. Pattinson N. R. Cholic acid binding by anionic glutathione-S-transferase from human liver cytosol. Biochem Biophys Res Commun. 1981 Sep 16;102(1):403–410. doi: 10.1016/0006-291x(81)91535-7. [DOI] [PubMed] [Google Scholar]
  28. Pearson W. R., Windle J. J., Morrow J. F., Benson A. M., Talalay P. Increased synthesis of glutathione S-transferases in response to anticarcinogenic antioxidants. Cloning and measurement of messenger RNA. J Biol Chem. 1983 Feb 10;258(3):2052–2062. [PubMed] [Google Scholar]
  29. Pickett C. B., Donohue A. M., Lu A. Y., Hales B. F. Rat liver glutathione S-transferase B: the functional mRNAs specific for the Ya Yc subunits are induced differentially by phenobarbital. Arch Biochem Biophys. 1982 May;215(2):539–543. doi: 10.1016/0003-9861(82)90113-8. [DOI] [PubMed] [Google Scholar]
  30. 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]
  31. Scully N. C., Mantle T. J. Tissue distribution and subunit structures of the multiple forms of glutathione S-transferase in the rat. Biochem J. 1981 Jan 1;193(1):367–370. doi: 10.1042/bj1930367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. 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]
  33. Singh S. V., Dao D. D., Partridge C. A., Theodore C., Srivastava S. K., Awasthi Y. C. Different forms of human liver glutathione S-transferases arise from dimeric combinations of at least four immunologically and functionally distinct subunits. Biochem J. 1985 Dec 15;232(3):781–790. doi: 10.1042/bj2320781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Smith G. J., Ohl V. S., Litwack G. Ligandin, the glutathione S-transferases, and chemically induced hepatocarcinogenesis: a review. Cancer Res. 1977 Jan;37(1):8–14. [PubMed] [Google Scholar]
  35. Smith G. J., Ohl V. S., Litwack G. Purification and properties of hamster liver ligandins, glutathione S-transferases. Cancer Res. 1980 Jun;40(6):1787–1790. [PubMed] [Google Scholar]
  36. Stockman P. K., Beckett G. J., Hayes J. D. Identification of a basic hybrid glutathione S-transferase from human liver. Glutathione S-transferase delta is composed of two distinct subunits (B1 and B2). Biochem J. 1985 Apr 15;227(2):457–465. doi: 10.1042/bj2270457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. 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]
  38. Tegtmeyer P., Schwartz M., Collins J. K., Rundell K. Regulation of tumor antigen synthesis by simain virus 40 gene A. J Virol. 1975 Jul;16(1):168–178. doi: 10.1128/jvi.16.1.168-178.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. 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]
  40. Tu C. P., Reddy C. C. On the multiplicity of rat liver glutathione S-transferases. J Biol Chem. 1985 Aug 25;260(18):9961–9964. [PubMed] [Google Scholar]
  41. Tu C. P., Weiss M. J., Li N. Q., Reddy C. C. Tissue-specific expression of the rat glutathione S-transferases. J Biol Chem. 1983 Apr 25;258(8):4659–4662. [PubMed] [Google Scholar]
  42. Tu C. P., Weiss M. J., Reddy C. C. Subunit composition of rat liver glutathione S-transferases. Biochem Biophys Res Commun. 1982 Sep 30;108(2):461–467. doi: 10.1016/0006-291x(82)90851-8. [DOI] [PubMed] [Google Scholar]
  43. Vander Jagt D. L., Hunsaker L. A., Garcia K. B., Royer R. E. Isolation and characterization of the multiple glutathione S-transferases from human liver. Evidence for unique heme-binding sites. J Biol Chem. 1985 Sep 25;260(21):11603–11610. [PubMed] [Google Scholar]

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