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.
Similar content being viewed by others
References
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.
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.
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.
Autrup, H. (2000). Genetic polymorphism in human xenobiotica metabolizing enzymes as susceptibility factors in toxic response. Mutation Research, 464(1), 65–76.
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.
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.
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.
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.
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.
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.
Hayes, J. D., Flanagan, J. U., & Jowsey, I. R. (2005). Glutathione transferases. Annual Review of Pharmacology and Toxicology, 45, 51–88.
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.
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.
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.
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.
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.
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.
Denizli, A. (2008). Heavy metal toxicology; applied molecular biology techniques in seafood (pp. 1–16). Erzurum, Turkey: Atatürk University Faculty of Agriculture.
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.
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.
Laemmli, D. K. (1970). Cleavage of structural proteins during in assembly of the head of bacteriophage T4. Nat, 227, 680–685.
Lineweaver, H., & Burk, D. (1934). The determination of enzyme dissociation constants. Journal of the American Chemical Society, 57, 658–666.
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
Priti, M., & Satish, S. (2014). Quail farming: an introduction. International Journal of Life Science, 2, 190–193.
Kahvecioğlu, Ö., Kartal, G., Güven, A., & Timur, S. (2004). Metallerin çevresel etkileri-III. İstanbul: GTÜ Metalürji ve Malzeme Mühendisliği Bölümü 15s.
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.
Jarup, L. (2003). Hazards of heavy metal contamination. British Medical Bulletin, 68, 167–182.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Loscalzo, J., & Freedman, J. (1986). Purification and characterization of human platelet glutathione-S transferase. Blood., 67(6), 1595–1599.
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.
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.
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.
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.
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.
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.
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.
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.
Author information
Authors and Affiliations
Corresponding author
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
About this article
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
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12668-020-00811-4