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. 1992 Jan 15;281(Pt 2):545–551. doi: 10.1042/bj2810545

Cloning and expression of a chick liver glutathione S-transferase CL 3 subunit with the use of a baculovirus expression system.

L H Chang 1, J Y Fan 1, L F Liu 1, S P Tsai 1, M F Tam 1
PMCID: PMC1130720  PMID: 1339283

Abstract

Glutathione S-transferase CL 3 subunits purified from 1-day-old-chick livers were digested with Achromobacter proteinase I and the resulting fragments were isolated for amino acid sequence analysis. An oligonucleotide probe was constructed accordingly for cDNA library screening. A cDNA clone of 1342 bases, pGCL301, encoding a protein of 26209 Da was isolated and sequenced. Including conservative substitutions, this protein has 75-79% sequence similarity to other Alpha family glutathione S-transferases. The coding sequence of pGCL301 was inserted into a baculovirus vector for infection of Spodoptera frugiperda (SF9) cells. The expressed protein has a high relative activity with ethacrynic acid (47% of the specific activity with 1-chloro-2,4-dinitrobenzene). The enzyme has a subunit molecular mass of 25.2 +/- 1.2 kDa (by SDS/PAGE), a pI of 9.45 and an absorption coefficient A1%1cm of 13.0 +/- 0.5 at 280 nm.

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Selected References

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

  1. Abramovitz M., Ishigaki S., Felix A. M., Listowsky I. Expression of an enzymatically active Yb3 glutathione S-transferase in Escherichia coli and identification of its natural form in rat brain. J Biol Chem. 1988 Nov 25;263(33):17627–17631. [PubMed] [Google Scholar]
  2. 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]
  3. 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]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. Chang L. H., Chuang L. F., Tsai C. P., Tu C. P., Tam M. F. Characterization of glutathione S-transferases from day-old chick livers. Biochemistry. 1990 Jan 23;29(3):744–750. doi: 10.1021/bi00455a022. [DOI] [PubMed] [Google Scholar]
  6. Chi H. C., Hsieh J. C., Hui C. F., Tam M. F. Modified method for double stranded DNA sequencing and synthetic oligonucleotide purification. Nucleic Acids Res. 1988 Nov 11;16(21):10382–10382. doi: 10.1093/nar/16.21.10382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chow N. W., Whang-Peng J., Kao-Shan C. S., Tam M. F., Lai H. C., Tu C. P. Human glutathione S-transferases. The Ha multigene family encodes products of different but overlapping substrate specificities. J Biol Chem. 1988 Sep 15;263(26):12797–12800. [PubMed] [Google Scholar]
  8. Coles B., Ketterer B. The role of glutathione and glutathione transferases in chemical carcinogenesis. Crit Rev Biochem Mol Biol. 1990;25(1):47–70. doi: 10.3109/10409239009090605. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. DeJong J. L., Chang C. M., Whang-Peng J., Knutsen T., Tu C. P. The human liver glutathione S-transferase gene superfamily: expression and chromosome mapping of an Hb subunit cDNA. Nucleic Acids Res. 1988 Sep 12;16(17):8541–8554. doi: 10.1093/nar/16.17.8541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gill S. C., von Hippel P. H. Calculation of protein extinction coefficients from amino acid sequence data. Anal Biochem. 1989 Nov 1;182(2):319–326. doi: 10.1016/0003-2697(89)90602-7. [DOI] [PubMed] [Google Scholar]
  12. Grove G., Zarlengo R. P., Timmerman K. P., Li N. Q., Tam M. F., Tu C. P. Characterization and heterospecific expression of cDNA clones of genes in the maize GSH S-transferase multigene family. Nucleic Acids Res. 1988 Jan 25;16(2):425–438. doi: 10.1093/nar/16.2.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Hewick R. M., Hunkapiller M. W., Hood L. E., Dreyer W. J. A gas-liquid solid phase peptide and protein sequenator. J Biol Chem. 1981 Aug 10;256(15):7990–7997. [PubMed] [Google Scholar]
  15. Hoesch R. M., Boyer T. D. Localization of a portion of the active site of two rat liver glutathione S-transferases using a photoaffinity label. J Biol Chem. 1989 Oct 25;264(30):17712–17717. [PubMed] [Google Scholar]
  16. Hsieh J. C., Liu L. F., Chen W. L., Tam M. F. Expression of Yb1 glutathione S-transferase using a baculovirus expression system. Biochem Biophys Res Commun. 1989 Aug 15;162(3):1147–1154. doi: 10.1016/0006-291x(89)90793-6. [DOI] [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. Kispert A., Meyer D. J., Lalor E., Coles B., Ketterer B. Purification and characterization of a labile rat glutathione transferase of the Mu class. Biochem J. 1989 Jun 15;260(3):789–793. doi: 10.1042/bj2600789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. 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]
  21. 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]
  22. 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]
  23. 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]
  24. 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]
  25. Moore R. E., Davies M. S., O'Connell K. M., Harding E. I., Wiegand R. C., Tiemeier D. C. Cloning and expression of a cDNA encoding a maize glutathione-S-transferase in E. coli. Nucleic Acids Res. 1986 Sep 25;14(18):7227–7235. doi: 10.1093/nar/14.18.7227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. 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]
  28. Ramanathan L., Guyer R. B., Karakawa W. W., Buss E. G., Clagett C. O. Immunochemistry of the glycopeptide derived from avian riboflavin-binding protein. Mol Immunol. 1979 Nov;16(11):935–941. doi: 10.1016/0161-5890(79)90094-4. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. 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]
  31. Tahir M. K., Guthenberg C., Mannervik B. Inhibitors for distinction of three types of human glutathione transferase. FEBS Lett. 1985 Feb 25;181(2):249–252. doi: 10.1016/0014-5793(85)80269-6. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. 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]
  34. 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]
  35. Wang R. W., Pickett C. B., Lu A. Y. Expression of a cDNA encoding a rat liver glutathione S-transferase Ya subunit in Escherichia coli. Arch Biochem Biophys. 1989 Mar;269(2):536–543. doi: 10.1016/0003-9861(89)90137-9. [DOI] [PubMed] [Google Scholar]
  36. Yeung T. C., Gidari A. S. Purification and properties of a chicken liver glutathione S-transferase. Arch Biochem Biophys. 1980 Dec;205(2):404–411. doi: 10.1016/0003-9861(80)90123-x. [DOI] [PubMed] [Google Scholar]

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