Skip to main content
SpringerLink
Log in

Glutathione S-transferase: Purification and Characterization from Quail (Coturnix coturnix japonica) Liver and the Impact of Some Metal Ions on Enzyme Activity

  • Published:
BioNanoScience Aims and scope Submit manuscript

Abstract

The objective of this study was to examine the inhibitory effect of some heavy metals on the glutathione S-transferase (GST) enzyme, which is one of the major enzymes of glutathione metabolism purified from quail liver tissues. The quail liver GST enzyme was purified with 15.86 EU/mg specific activity, in a yield of 12.36% and 46.1 purification fold by ammonium sulfate precipitation and glutathione-agarose affinity chromatography. The molecular weight of subunits of the enzyme and the purity were determined by SDS-PAGE. In the characterization studies, the optimum pH of the GST enzyme was determined to be pH = 8.0 in Tris/HCl buffer. Optimum ionic strength was determined to be 140 mM in Tris/HCI buffer. Stable pH was found to be pH = 8.5 in Tris/HCl buffer. Optimum temperature was found to be 50 °C. KM and Vmax values for substrates 1-chloro-2,4-dinitrobenzene (CDNB) and GSH of the enzyme were determined to be KM 0.048 mM Vmax 0.479 EU/mL and KM 0.114 mM Vmax 0.672 EU/mL, respectively. In vitro inhibition effects of metal ions, including Ag+, Ni2+, Cd2+, Fe2+, Pb2+, Co2+ Zn2+, and Al3+, were investigated on the GST enzyme activity. The results showed that Ag+, Cd2+, Ni2+ Zn2+, and Al3+ metal ions inhibited GST enzyme (IC50 values 0.239, 0.250, 0.265, 0.320, 0.594 mM, respectively), while Fe2+, Pb2+, and Co2+ metal ions activated the enzyme. Finally, Ki values and inhibition types for these substances were determined by Lineweaver-Burk graphs.

Graphical abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Gülçin, İ., Scozzafava, A., Supuran, C. T., Akıncıoğlu, H., Koksal, Z., Turkan, F., & Alwasel, S. (2016). The effect of caffeic acid phenethyl ester (CAPE) on metabolic enzymes including acetylcholinesterase, butyrylcholinesterase, glutathione S-transferase, lactoperoxidase, and carbonic anhydrase isoenzymes I, II, IX, and XII. Journal of Enzyme Inhibition and Medicinal Chemistry, 31(6), 1095–1101.

    Google Scholar 

  2. Temel, Y., Koçyigit, U. M., Taysı, M. Ş., Gökalp, F., Gürdere, M. B., Budak, Y., Gulcin, I., & Çiftci, M. (2018). Purification of glutathione S-transferase enzyme from quail liver tissue and inhibition effects of (3aR, 4S, 7R, 7aS)-2-(4-((E)-3-(aryl) acryloyl) phenyl)-3a, 4, 7, 7a-tetrahydro-1H-4, 7-methanoisoindole-1, 3 (2H)-dione derivatives on the enzyme activity. Journal of Biochemical and Molecular Toxicology, 32(3), e22034.

    Google Scholar 

  3. Sheehan, D., Meade, G., Foley, V. M., & Dowd, C. A. (2001). Structure, function and evolution of glutathione transferases: implications for classification of non-mammalian members of an ancient enzyme superfamily. The Biochemical Journal, 360(1), 1–16.

    Google Scholar 

  4. Autrup, H. (2000). Genetic polymorphism in human xenobiotica metabolizing enzymes as susceptibility factors in toxic response. Mutation Research, 464(1), 65–76.

    Google Scholar 

  5. Işik, M., Demir, Y., Kirici, M., Demir, R., Şimşek, F., & Beydemir, Ş. (2015). Changes in the anti-oxidant system in adult epilepsy patients receiving anti-epileptic drugs. Archives of Physiology and Biochemistry, 121, 97–102.

    Google Scholar 

  6. Türkan, F., Huyut, Z., Demir, Y., Ertaş, F., & Beydemir, Ş. (2019). The effects of some cephalosporins on acetylcholinesterase and glutathione S-transferase: an in vivo and in vitro study. Archives of Physiology and Biochemistry, 125, 235–243.

    Google Scholar 

  7. Özaslan, M. S., Demir, Y., Küfrevioğlu, O. I., & Çiftci, M. (2017). Some metals inhibit the glutathione S-transferase from Van Lake fish gills. Journal of Biochemical and Molecular Toxicology, 31(11), e21967.

    Google Scholar 

  8. Özaslan, M. S., Demir, Y., Aslan, H. E., Beydemir, Ş., & Küfrevioğlu, Ö. İ. (2018). Evaluation of chalcones as inhibitors of glutathione S-transferase. Journal of Biochemical and Molecular Toxicology, 32, 1–6.

    Google Scholar 

  9. Özaslan, M. S., Demir, Y., Aksoy, M., Küfrevioğlu, Ö. I., & Beydemir, Ş. (2018). Inhibition effects of pesticides on glutathione-S-transferase enzyme activity of Van Lake fish liver. Journal of Biochemical and Molecular Toxicology, 32(9), e22196.

    Google Scholar 

  10. Ceylan, H., Demir, Y., & Beydemir, Ş. (2019). Inhibitory effects of usnic and carnosic acid on some metabolic enzymes: an in vitro study. Protein and Peptide Letters, 26(5), 364–370.

    Google Scholar 

  11. Hayes, J. D., Flanagan, J. U., & Jowsey, I. R. (2005). Glutathione transferases. Annual Review of Pharmacology and Toxicology, 45, 51–88.

    Google Scholar 

  12. Bucciarelli, T., Sacchetta, P., Pennelli, A., Cornelio, L., Romagnoli, R., Melino, S., & Di Ilio, C. (1999). Characterization of toad liver glutathione transferase. Biochimica et Biophysica Acta, 1431(1), 189–198.

    Google Scholar 

  13. Caglayan, C., Taslimi, P., Türk, C., Kandemir, F. M., Demir, Y., & Gulcin, İ. (2019). Purification and characterization of the carbonic anhydrase enzyme from horse mackerel (Trachurus trachurus) muscle and the impact of some metal ions and pesticides on enzyme activity. Comparative Biochemistry and Physiology, Part C: Toxicology & Pharmacology, 226, 108605.

    Google Scholar 

  14. Caglayan, C., Taslimi, P., Türk, C., Gulcin, İ., Kandemir, F. M., Demir, Y., & Beydemir, Ş. (2020). Inhibition effects of some pesticides and heavy metals on carbonic anhydrase enzyme activity purified from horse mackerel (Trachurus trachurus) gill tissues. Environmental Science and Pollution Research, 27, 10607–10616.

    Google Scholar 

  15. Kirici, M., Kirici, M., Demir, Y., Beydemir, S., & Atamanalp, M. (2016). The effect of AL3+ and Hg4+ on glucose 6-phosphate dehydrogenase from capoeta umbla kidney. Applied Ecology and Environmental Research, 14, 253–264.

    Google Scholar 

  16. Donkın, S. G., & Ohlson, D. L. (2000). Teaf, C.M. Principle of toxicology, environmental and industrial applications. In P. L. Williams, R. C. James, & S. M. Roberts (Eds.), (2nd ed., p. 325). New York: Wiley ABD.

    Google Scholar 

  17. Goyer, R. A., & Clarkson, T. W. (2001). Toxic Effects of Metals. In C. D. Klaassen (Ed.), Casarette & Doull’s Toxicology-the Basic Science of Poisons (6th ed., pp. 827–844). New York: McGraw-Hill. ABD.

    Google Scholar 

  18. Denizli, A. (2008). Heavy metal toxicology; applied molecular biology techniques in seafood (pp. 1–16). Erzurum, Turkey: Atatürk University Faculty of Agriculture.

    Google Scholar 

  19. Habig, W. H., Pabst, M. J., & Jakoby, W. B. (1974). Glutathione S-transferase A. A novel kinetic mechanism in which the major reaction pathway depends on substrate concentration. The Journal of Biological Chemistry, 249, 7140–7150.

    Google Scholar 

  20. Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248–251.

    Google Scholar 

  21. Laemmli, D. K. (1970). Cleavage of structural proteins during in assembly of the head of bacteriophage T4. Nat, 227, 680–685.

    Google Scholar 

  22. Lineweaver, H., & Burk, D. (1934). The determination of enzyme dissociation constants. Journal of the American Chemical Society, 57, 658–666.

    Google Scholar 

  23. Zhou, Z., Tian, W., Wang, J., Liu, H., Cao, L. (2012) Food consumption trends in China 2012. http://www.agriculture.gov.au/. Accessed 25 Dec 2018

  24. Priti, M., & Satish, S. (2014). Quail farming: an introduction. International Journal of Life Science, 2, 190–193.

    Google Scholar 

  25. Kahvecioğlu, Ö., Kartal, G., Güven, A., & Timur, S. (2004). Metallerin çevresel etkileri-III. İstanbul: GTÜ Metalürji ve Malzeme Mühendisliği Bölümü 15s.

    Google Scholar 

  26. Temel, Y., Küfrevioğlu, Ö. İ., & Ciftci, M. (2017). Investigation of the effects of purification and characterization of Turkey (Meleagris gallopavo) liver mitochondrial thioredoxin reductase enzyme and some metal ions on enzyme activity. Turkish Journal of Chemistry, 41(1), 48–60.

    Google Scholar 

  27. Jarup, L. (2003). Hazards of heavy metal contamination. British Medical Bulletin, 68, 167–182.

    Google Scholar 

  28. Gupta, U. C., & Gupta, S. C. (1998). Trace elements toxicity relationships to crop production and livestock and human health: implication for management. Communications in Soil Science and Plant Analysis, 29, 1491–1522.

    Google Scholar 

  29. Innocenti, A., Öztürk Sarıkaya, S. B., Gülçin, İ., & Supuran, C. T. (2010). Carbonic anhydrase inhibitors. Inhibition of mammalian isoforms I–XIV with a series of natural product polyphenols and phenolic acids. Bioorganic & Medicinal Chemistry, 18(6), 2159–2164.

    Google Scholar 

  30. Öztürk Sarıkaya, S. B., Gülçin, İ., & Supuran, C. T. (2010). Carbonic anhydrase inhibitors, inhibition of human erythrocyte isozymes I and II with a series of phenolic acids. Chemical Biology & Drug Design, 75, 515–520.

    Google Scholar 

  31. Temel, Y., & Kocyigit, U. M. (2017). Purification of glucose-6-phosphate dehydrogenase from rat (Rattus norvegicus) erythrocytes and inhibition effects of some metal ions on enzyme activity. Journal of Biochemical and Molecular Toxicology, 31, e21927.

    Google Scholar 

  32. Ceylan, M., Kocyigit, U. M., Usta, N. C., Gürbüzlü, B., Temel, Y., Alwasel, S. H., & Gülçin, İ. (2017). Synthesis, carbonic anhydrase I and II isoenzymes inhibition properties, and antibacterial activities of novel tetralone-based 1, 4-benzothiazepine derivatives. Journal of Biochemical and Molecular Toxicology, 31, e21872.

    Google Scholar 

  33. Bayındır, S., Ayna, A., Temel, Y., & Ciftci, M. (2018). The synthesis of new oxindoles as analogs of natural product 3,3′-bis(indolyl)oxindole and in vitro evaluation for enzyme activity of G6PD and 6PGD. Turkish Journal of Chemistry, 42(2), 332–345.

    Google Scholar 

  34. Bayindir, S., Temel, Y., Ayna, A., & Ciftci, M. (2018). The synthesis of N-benzoylindoles as inhibitors of rat erythrocyte glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase. Journal of Biochemical and Molecular Toxicology, 32, e22193.

    Google Scholar 

  35. Tugrak, M., Gul, H. I., Sakagami, H., Gulcin, I., & Supuran, C. T. (2018). New azafluorenones with cytotoxic and carbonic anhydrase inhibitory properties: 2-Aryl-4-(4-hydroxyphenyl)-5H-indeno [1, 2-b] pyridin-5-ones. Bioorganic Chemistry, 81, 433–439.

    Google Scholar 

  36. Gulçin, İ., Taslimi, P., Aygün, A., Sadeghian, N., Bastem, E., Kufrevioglu, O. I., & Şen, F. (2018). Antidiabetic and antiparasitic potentials: Inhibition effects of some natural antioxidant compounds on α-glycosidase, α-amylase and human glutathione S-transferase enzymes. International Journal of Biological Macromolecules, 119, 741–746.

    Google Scholar 

  37. Taslimi, P., Aslan, H. E., Demir, Y., Oztaskin, N., Maraş, A., Gulçin, İ., & Goksu, S. (2018). Diarylmethanon, bromophenol and diarylmethane compounds: Discovery of potent aldose reductase, α-amylase and α-glycosidase inhibitors as new therapeutic approach in diabetes and functional hyperglycemia. International Journal of Biological Macromolecules, 119, 857–863.

    Google Scholar 

  38. Loscalzo, J., & Freedman, J. (1986). Purification and characterization of human platelet glutathione-S transferase. Blood., 67(6), 1595–1599.

    Google Scholar 

  39. Young, P. R., & Briedis, A. V. (1989). Purification and kinetic mechanism of the major glutathione S-transferase from bovine brain. The Biochemical Journal, 257(2), 541–548.

    Google Scholar 

  40. Awasthi, Y. C., Doa, D. D., & Saneto, R. P. (1980). İnterrelationship between anionic and cationic forms of glutathione S-transferases of human liver. The Biochemical Journal, 191, 1–10.

    Google Scholar 

  41. Huang, Q., Liang, L., Wei, T., Zhang, D., & Zeng, Q. Y. (2008). Purification and partial characterization of glutathione transferase from the teleost Monopterus albus. Comparative Biochemistry and Physiology - Part C, 147(1), 96–100.

    Google Scholar 

  42. Erat, M., & Şakiroğlu, H. (2013). The effect of some antineoplastic agents on glutathione S-transferase from human eriytrocytes. Journal of Enzyme Inhibition and Medicinal Chemistry, 28(4), 711–716.

    Google Scholar 

  43. Comakli, V., Kuzu, M., & Demirdag, M. (2015). Characterization and purification of Glutathione S-transferase from the liver and gill tissues of Ağrı Balık Lake trout Salmo trutta labrax and the effects of heavy metal ions on its activity. Journal of Aquatic Animal Health, 27, 145–151.

    Google Scholar 

  44. Kucuk, M., & Gulcin, İ. (2016). Purification and characterization of the carbonic anhydrase enzyme from Black Sea trout (Salmo trutta Labrax Coruhensis) kidney and inhibition effects of some metal ions on enzyme activity. Environmental Toxicology and Pharmacology, 44, 134–139.

    Google Scholar 

  45. Güvercin, S., Erat, M., & Şakiroğlu, H. (2008). Determination of some kinetic and characteristic properties of glutathione S-transferase from bovine erythrocytes. Protein and Peptide Letters, 15, 6–12.

    Google Scholar 

  46. Toribio, F., Martinez-Lara, E., Pascual, P., & Lopez-Barea, J. (1996). Methods for purification of glutathione peroxidase and related enzymes. Journal of Chromatography B, 684(1–2), 77–97.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Chemistry, Arts and Science Faculty, Bingol University, Bingol, Turkey

    Mehmet Şerif Taysi

  2. Solhan Health Services Vocational School, Bingol University, 12000, Bingol, Turkey

    Yusuf Temel

Authors
  1. Mehmet Şerif Taysi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yusuf Temel
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yusuf Temel.

Ethics declarations

Conflict of Interest

None

Research Involving Humans and Animals Statement

This procedure was carried out with approval from the Bingol University Local Ethics Committee for Animal Experiments (BUHADYEK-29/06/2018-12212).

Informed Consent

None.

Funding Statement

The authors thank the Research Fund of Bingol University for funding the project (BAP-SSHMYO.2017.00.001).

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Highlights

• Glutathione S-transferase enzyme was purified from the quail (Coturnix coturnix japonica) liver.

• GST enzyme was characterized.

• SDS-PAGE was performed for enzyme purity and subunit.

• The effects of some heavy metals ions were determined on GST enzyme activity.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Taysi, M.Ş., Temel, Y. Glutathione S-transferase: Purification and Characterization from Quail (Coturnix coturnix japonica) Liver and the Impact of Some Metal Ions on Enzyme Activity. BioNanoSci. 11, 91–98 (2021). https://doi.org/10.1007/s12668-020-00811-4

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12668-020-00811-4

Keywords

Search

Navigation