Purification of Glutathione Reductase From Some Tissues of Capoeta umbla and the Inhibitory Effects of Some Metal Ions on Enzyme Activity

Year 2021, Volume: 10 Issue: 2, 193 - 200, 05.06.2021

Muammer Kırıcı , Mahinur Kırıcı , Muhammed Atamanalp , Şükrü Beydemir

https://doi.org/10.33714/masteb.769454

Cited By: 3

Abstract

The aim of this study was to determine the in vitro inhibitory effects of some metal ions (silver ion (Ag⁺), cadmium ion (Cd²⁺), cobalt ion (Co²⁺), copper ion (Cu²⁺), nickel ion (Ni²⁺), lead ion (Pb²⁺) and zinc ion (Zn²⁺)) on glutathione reductase (GR) enzyme activities that purified from the gill, kidney and liver tissues of Capoeta umbla. For this purpose, the enzyme was purified from the gill, kidney and liver of C. umbla freshwater fish using ammonium sulfate precipitation and affinity column chromatography methods using 2′,5′-ADP Sepharose 4B. Within this study, the GR enzyme was purified for the first time from the tissues of C. umbla. Enzyme purity and molecular weight were determined using the sodium dodecyl sulfate polyacrylamide gel electrophoresis method. In addition, the inhibitory effects of different metal ions (Ag⁺, Cd²⁺, Co²⁺, Cu²⁺, Ni²⁺, Pb²⁺ and Zn²⁺) on GR enzyme activities of the gill, kidney and liver tissue of C. umbla were investigated under in vitro conditions. The metal ion concentrations inhibiting 50% of enzyme activity (IC₅₀) were obtained by plotting activity percentage versus [I] figures. Finally, the dissociation constants of the enzyme inhibitor complex (Ki), and the inhibition types, were calculated from Lineweaver–Burk plots. In vitro inhibition rank order was determined as Ag⁺>Co²⁺>Pb²⁺>Zn²⁺>Cu²⁺ for C. umbla gill GR; Ag⁺>Pb²⁺>Co²⁺> Ni²⁺>Zn²⁺ for C. umbla liver GR; Ag⁺>Cu²⁺>Co²⁺>Pb²⁺>Ni²⁺ for C. umbla kidney GR. From these results, we showed that Ag⁺ metal ion is the most potent inhibitor of GR enzyme on gill, liver and kidney tissues. Our results also demonstrate that these metals might be dangerous at low micromolar concentrations for C. umbla GR enzyme.

Keywords

Glutathione reductase, Metal toxicity, Capoeta umbla, Enzyme inhibition

Supporting Institution

-

Project Number

-

References

• Akkemik, E., Senturk, M., Ozgeris, F. B., Taser, P. & Ciftci, M. (2011). In vitro effects of some drugs on human erythrocyte glutathione reductase. Turkish Journal of Medical Sciences, 41(2): 235–241. https://doi.org/10.3906/sag-1002-4
• Altun, M., Türkoğlu V. & Çelik, İ. (2015). The effect of some antibiotics on glutathione reductase enzyme purified from liver and erythrocyte of Lake Van pearl mullet. Pharmaceutical Biology, 53(11): 1647-1652. https://doi.org/10.3109/13880209.2014.997830
• 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(1-2): 248–254. https://doi.org/10.1016/0003-2697(76)90527-3
• Carlberg, I. & Mannervik, B. (1975). Purification and characterization of the flavoenzyme glutathione reductase from rat liver. Journal of Biological Chemistry, 250(14): 5475–5480.
• Carlberg, I., Altmejd, B. & Mannervik, B. (1981). Purification and immunological studies of glutathione reductase from rat liver. Evidence for an antigenic determinant at the nucleotide-binding domain of the enzyme. Biochimica et Biophysica Acta, 677(1): 146–152. https://doi.org/10.1016/0304-4165(81)90156-2
• Ceyhun, S. B., Senturk, M., Yerlikaya, E., Erdoğan, O., Kufrevioglu, I. O. & Ekinci, D. (2011). Purification and characterization of carbonic anhydrase from the teleost fish Dicentrarchus labrax (European seabass) liver and toxicological effects of metals on enzyme activity. Environmental Toxicology and Pharmacology, 32(1): 69-74. https://doi.org/10.1016/j.etap.2011.03.013
• Ekinci, D. & Beydemir, S. (2010). Risk assessment of pesticides and fungicides foracid-base regulation and salt transport in rainbow trout tissues. Pesticide Biochemistry and Physiology, 97(1): 66–70. https://doi.org/10.1016/j.pestbp.2009.12.006
• Fraternale, A., Paoletti, M. F., Casabianca, A., Nencioni, L., Garaci, E., Palamara, A. T. & Magnani, M. (2009). GSH and analogs in antiviral therapy. Molecular Aspects of Medicine, 30(1-2): 99–110. https://doi.org/10.1016/j.mam.2008.09.001
• Gironi, M., Bianchi, A., Russo, A., Alberoni, M., Ceresa, L., Angelini, A., Cursano, C., Mariani, E., Nemni, R., Kullmann, C., Farina, E. & Boneschi, F. M. (2011). Oxidative imbalance in different neurodegenerative diseases with memory impairment. Neurodegenerative Diseases, 8(3): 129–137. https://doi.org/10.1159/000319452
• Innocenti, A., Ozturk Sarikaya, S. B., Gulcin, I. & Supuran, C. T. (2010). Carbonic anhydrase inhibitors. Inhibition of mammalian isoforms I–XIV with a series of natural product polyphenols and phenolic acids. Bioorganic and Medicinal Chemistry, 18(6): 2159–2164. https://doi.org/10.1016/j.bmc.2010.01.076
• Kirici, M., Atamanalp, M., Kirici, M. & Beydemir, S. (2017a). In vitro effects of some metal ions on glutathione reductase in the gills and liver of Capoeta trutta. Regulatory Mechanisms in Biosystems, 8(1): 66–70. https://doi.org/10.15421/021712
• Kırıcı, M., Atamanalp, M., Kırıcı, M. & Beydemir, Ş. (2020). Purification of glucose 6-phosphate dehydrogenase from Capoeta umbla gill and liver tissues and inhibition effects of some metal ions on enzyme activity. Marine Science and Technology Bulletin, 9(2): 92-101. https://doi.org/10.33714/masteb.709377
• Kırıcı, M., Kırıcı, M. & Atamanalp, M. (2017c). In vitro inhibition effects of some metal ions on glutathione reductase purified from Capoeta trutta kidney. Aquaculture Studies, 17(4): 385-394. http://doi.org/10.17693/yunusae.v17i31121.337640
• Kirici, M., Kirici, M., Beydemir, S. & Atamanalp, M. (2016a). Purification of carbonic anhydrase from Capoeta umbla (Heckel, 1843) gills and toxicological effects of some metals on enzyme activity. Turkish Journal of Fisheries and Aquatic Sciences, 16(1): 169–175. https://doi.org/10.4194/1303-2712-v16_1_17
• Kirici, M., Kirici, M., Beydemir, S. & Bulbul, M., (2017b). Purification of glucose 6-phosphate dehydrogenase from gilthead sea bream (Sparus aurata) gill and liver tissues and inhibition effects of some metal ions on enzyme activity. Fresenius Environmental Bulletin, 26(2): 7074-7082.
• Kirici, M., Kirici, M., Demir, Y., Beydemir, S. & Atamanalp, M. (2016b). The effect of Al3+ and Hg2+ on glucose 6-phosphate dehydrogenase from Capoeta umbla kidney. Applied Ecology and Environmental Research, 14(2): 253-264. http://dx.doi.org/10.15666/aeer/1402_253264
• Kırıcı, M., Kırıcı, M., Işık, M. & Atamanalp, M. (2015). İmidacloprid ve Lambda-cyhalothrin'in Capoeta capoeta umbla böbrek dokusunda glikoz 6-fosfat dehidrogenaz enzimi üzerine in vitro etkileri. Türkiye Tarımsal Araştırmalar Dergisi, 2(1): 8-14. https://doi.org/10.19159/tutad.41219
• Knapen, M. F. C. M., Zusterzeel, P. L. M., Peters, W. H. M. & Steegers, E. A. P. (1999). Glutathione and glutathione-related enzymes in reproduction. European Journal of Obstetrics and Gynecology and Reproductive Biology, 82(2): 171–184. https://doi.org/10.1016/S0301-2115(98)00242-5
• Kucuk, M. & Gulcin, I. (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. https://doi.org/10.1016/j.etap.2016.04.011
• Kuzu, M., Aslan, A., Ahmed, I., Comakli, V., Demirdag, R. & Uzun, N. (2016). Purification of glucose-6-phosphate dehydrogenase and glutathione reductase enzymes from the gill tissue of Lake Van fish and analyzing the effects of some chalcone derivatives on enzyme activities. Fish Physiology and Biochemistry, 42(2): 483-491. https://doi.org/10.1007/s10695-015-0153-7
• Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227: 680–685.
• Le Trang, N., Bhargava, K. K. & Cerami, A. (1983). Purification of glutathione reductase from gerbil liver in two steps. Analytical Biochemistry, 133(1): 94–99. https://doi.org/10.1016/0003-2697(83)90226-9
• Lineweaver, H. & Burk, D. (1934). The determination of enzyme dissociation constants. Journal of the American Chemical Society, 56(3): 685-666. https://doi.org/10.1021/ja01318a036
• Loro, V. L., Jorge, M. B., da Silva, K. R. & Wood, C. M. (2012). Oxidative stress parameters and antioxidant response to sublethal waterborne zinc in a euryhaline teleost Fundulus heteroclitus: protective effects of salinity. Aquatic Toxicology, 110-111: 187–193. https://doi.org/10.1016/j.aquatox.2012.01.012
• Ozaslan, M. S., Demir, Y., Kufrevioglu, O. I. & Ciftci, M. (2017). Some metals inhibit the glutathione S-transferase from Van Lake fish gills. Journal of Biochemical and Molecular Toxicology, 31(11): e21967. https://doi.org/10.1002/jbt.21967
• Qu, R., Feng, M., Wang, X., Qin, L., Wang, C., Wang, Z. & Wang, L. (2014). Metal accumulation and oxidative stress biomarkers in liver of freshwater fish Carassius auratus following in vivo exposure to waterborne zinc under different pH values. Aquatic Toxicology, 150: 9–16. https://doi.org/10.1016/j.aquatox.2014.02.008
• Raza, H. (2011). Dual localization of glutathione S-transferase in the cytosol and mitochondria: implications in oxidative stress, toxicity and disease. Federation of European Biochemical Societies Journal, 278(22): 4243–4251. https://doi.org/10.1111/j.1742-4658.2011.08358.x
• Schirmer, R. H., Krauth-Siegel, R. L. & Schulz, G. E. (1989). Coenzymes and cofactors (pp. 553–596). In: Dolphin, D., Poulson, R. & Avramovic, O. (Eds.), Glutathione. New York, USA: John Wiley and Sons.
• Simic, T., Savic-Radojevic, A., Pljesa-Ercegovac, M., Matic, M. & Mimic-Oka, J. (2009). Glutathione S-transferases in kidney and urinary bladder tumors. Nature Reviews Urology, 6(5): 281–289. https://doi.org/10.1038/nrurol.2009.49
• Taser, P. & Ciftci, M. (2012). Purification and characterization of glutathione reductase from turkey liver. Turkish Journal of Veterinary and Animal Sciences, 36(5): 546–553. https://doi.org/10.3906/vet-1103-5
• Tchaikovskaya, T., Fraifeld, V., Urphanishvili, T., Andorfer, J. H., Davies, P. & Listowsky, I. (2005). Glutathione S-transferase hGSTM3 and ageing-associated neurodegeneration: relationship to Alzheimer’s disease. Mechanisms of Ageing and Development, 126(2): 309–315. https://doi.org/10.1016/j.mad.2004.08.029
• Tekman, B., Ozdemir, H., Senturk, M. & Ciftci, M. (2008). Purification and characterization of glutathione reductase from rainbow trout (Oncorhynchus mykiss) liver and inhibition effects of metal ions on enzyme activity. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 148(2): 117–121. https://doi.org/10.1016/j.cbpc.2008.04.005
• Yadav, S. S., Srikanth, E., Singh, N. & Rathaur, S. (2013). Identification of glutathione reductase and TrxR systems in Setaria cervi: Purification and characterization of glutathione reductase. Parasitology International, 62(2): 193–198. https://doi.org/10.1016/j.parint.2012.12.008