Purification of Glucose 6-Phosphate Dehydrogenase from Capoeta umbla Gill and Liver Tissues and Inhibition Effects of Some Metal Ions on Enzyme Activity

Abstract

In this study, the in vitro effects of some metal ions (Ag⁺, Cd²⁺, Cu²⁺, Fe²⁺, Ni²+ and Pb²⁺) on freshwater fish Capoeta umbla liver and gill glucose 6-phosphate dehydrogenase (G6PD) have been investigated. For this purpose, C. umbla liver and gill G6PD enzymes were purified, with a specific activity of 31.52 and 22.83 EU/mg protein, 3353.2 and 736.5 fold in a yield of 19.28% and 23.15%, respectively. In order to control the enzyme purification SDS-PAGE showed a single band for the enzyme. In addition, in vitro effects of metal ions on enzyme activity were researched. As a result, Ag⁺, Cd²⁺, Cu²⁺, Ni²⁺ and Pb²⁺ inhibited fish liver G6PD; Ag⁺, Cd²⁺, Fe²⁺, Ni²+ and Pb²+ inhibited fish gill G6PD. Besides, it was found that the most effective inhibitor of G6PD enzyme within metal ions used was Ag⁺. Our results also demonstrate that these metals might be dangerous at low micromolar concentrations for fish G6PD enzyme.

Keywords

• Glucose 6-phosphate dehydrogenase,Capoeta umbla,Enzyme purification,Inhibition,Metal ions,Liver

References

1. Adem, S. & Ciftci, M. (2012). Purification of rat kidney glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and glutathione reductase enzymes using 2′, 5′-ADP Sepharose 4B affinity in a single chromatography step. Protein Expression and Purification, 81(1): 1–4. https://doi.org/10.1016/j.pep. 2011.08.031
2. Adem, S. & Ciftci, M. (2016). Purification and characterization of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and glutathione reductase from rat heart and inhibition effects of furosemide, digoxin, and dopamine on the enzymes activities. Journal of Biochemical and Molecular Toxicology, 30(6): 295-301. https://doi.org/10.1002/ jbt.21793
3. Alici, H. A., Ekinci, D. & Beydemir, S. (2008). Intravenous anesthetics inhibit human paraoxonase-1 (PON1) activity in vitro and in vivo. Clinical Biochemistry, 41(16-17): 1384-1390. https://doi.org/10.1016/j.clinbiochem. 2008.06.017
4. Alım, Z. & Beydemir, Ş. (2012). Effects of some anti-neoplastic drugs on sheep liver sorbitol dehydrogenase. Archives of Physiology and Biochemistry, 118(5): 244-252. https://doi.org/10.3109/13813455.2012.688055
5. Atli, G., Alptekin, O., Tukel, S. & Canli, M. (2006). Response of catalase activity to Ag+, Cd2+, Cr6+, Cu2+ and Zn2+ in five tissues of freshwater fish Oreochromis niloticus. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 143(2): 218-224. https://doi.org/10.1016/j.cbpc.2006.02.003
6. Bautista, J. M., Garrido-Pertierra, A. & Soler, G. (1988). Glucose-6-phosphate dehydrogenase from Dicentrarchus labrax liver: kinetic mechanism and kinetics of NADPH inhibition. Biochimica et Biophysica Acta, 967(3): 354–363. https://doi.org/10.1016/0304-4165(88)90098-0
7. Beutler, E. (1971). Red Cell Metabolism Manual of Biochemical Methods. London, UK: Academic Press. 68p.
8. Beutler, E. (2008). Glucose-6-phosphate dehydrogenase deficiency: a historical perspective. Blood, 111(1): 16-24. https://doi.org/10.1182/blood-2007-04-077412

9. Beydemir, Ş., Yılmaz, H., Çiftçi, M., Bakan, E. & Küfrevioğlu, Ö. İ. (2003). Purification of glucose 6-phosphate dehydrogenase from goose erythrocytes and kinetic properties. Turkish Journal of Veterinary and Animal Sciences, 27(5): 1179–1185.
10. Bradford, M. M. A. (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
11. Caglayan, C., Taslimi, P., Türk, C., Kandemir, M. F., Demir, Y. & Gulcin, I. (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. https://doi.org/10.1016/j.cbpc.2019.108605
12. Cankaya, M., Sisecioglu, M., Ciftci, M. & Ozdemir, H. (2011). Effect of some metals on metal ions on trout liver G6PD. Reserch Journal of Environmental Toxicology, 5(6): 385-391. https://doi.org/10.3923/rjet.2011.385.391
13. Cerqueira, C. C. C. & Fernandes, M. N. (2002). Gill tissue recovery after copper exposure and blood parameter responses in the tropical fish, Prochilodus scrofa. Ecotoxicology and Environmental Safety, 52(2): 83-91. https://doi.org/10.1006/eesa.2002.2164
14. Çoban, M. Z., Gündüz F., Demirol, F., Örnekçi, G. N., Karakaya, G., Türkgülü, İ. & Alp, A. (2013). Population dynamics and stock assessment of Capoeta umbla (Heckel, 1843) in Lake Hazar, Elazığ, Turkey. Turkish Journal of Fisheries and Aquatic Sciences, 13(2): 221-231. https://doi.org/10.4194/1303-2712-v13_2_04
15. Comakli, V., Akkemik, E., Ciftci, M. & Kufrevioglu, O. I. (2015). Purification and characterization of glucose 6-phosphate dehydrogenase enzyme from rainbow trout (Oncorhynchus mykiss) liver and investigation of the effects of some metal ions on enzyme activity. Toxicology and Industrial Health, 31(5): 403-411. https://doi.org/10.1177/0748233713475514
16. Demir, Y., Oruç, E. & Topal, A. (2016). Carbonic anhydrase activity responses and histopathological changes in gill and liver tissues after acute exposure to chromium in brown trout juveniles. Hacettepe Journal of Biology and Chemistry, 44(4): 515-523. https://doi.org/10.15671/ HJBC.2016.132
17. Ekinci, D. & Beydemir, Ş. (2009). Evaluation of the impacts of antibiotic drugs on PON 1; a major bioscavenger against cardiovascular diseases. European Journal of Pharmacology, 617(1-3): 84-89. https://doi.org/10.1016/ j.ejphar.2009.06.048
18. Ekinci, D. & Beydemir, Ş. (2010). Risk assessment of pesticides and fungicides for acid-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
19. Ekinci, D., Beydemir, Ş. & Küfrevioğlu, Ö. İ. (2007). In vitro inhibitory effects of some heavy metals on human erythrocyte carbonic anhydrases. Journal of Enzyme Inhibition and Medicinal Chemistry, 22(6): 745-750. https://doi.org/10.1080/14756360601176048
20. Heath, A. G. (1987). Water Pollution and Fish Physiology. Florida, USA: CRC Press, 245p.
21. Hopa, E., Sinan, S. & Sevindik, E. (2015). The effects of deltamethrin and diazinon chemicals on human erythrocyte glucose-6-phosphate dehydrogenase enzyme. Journal of Applied Biological Sciences, 9(1): 31-35.
22. Hu, W., Zhi, L., Zhuo, M. Q., Zhu, Q. L., Zheng, J. L., Chen, Q. L., Gong, Y. & Liu, C. X. (2013). Purification and characterization of glucose 6-phosphate dehydrogenase (G6PD) from grass carp (Ctenopharyngodon idella) and inhibition effects of several metal ions on G6PD activity in vitro. Fish Physiology and Biochemistry, 39: 637-647. https://doi.org/10.1007/s10695-012-9726-x
23. Hunsaker, E. W & Franz, K. J. (2019). Emerging opportunities to manipulate metal trafficking for therapeutic benefit. Inorganic Chemistry, 58(20): 13528-13545. https://doi.org/10.1021/acs.inorgchem.9b01029
24. Jomova, K. & Valko, M. (2011). Advances in metal-induced oxidative stress and human disease. Toxicology, 283(2-3): 65-87. https://doi.org/10.1016/j.tox.2011.03.001
25. Kaya, E. D. Söyüt, H. & Beydemir, Ş. (2013). Carbonic anhydrase activity from the gilthead sea bream (Sparus aurata) liver: The toxicological effects of heavy metals. Environmental Toxicology and Pharmacology, 36(2): 514-521. https://doi.org/10.1016/j.etap.2013.05.019
26. Kirici, M., Atamanalp, M., Kirici, M. & Beydemir, Ş. (2017b). 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
27. Kirici, M., Kirici, M., Beydemir, S. & Atamanalp, M. (2016b). 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
28. Kirici, M., Kirici, M., Beydemir, S. & Bulbul, M. (2016c). In vitro toxicity effects of some insecticides on gilthead sea bream (Sparus aurata) liver glucose 6-phosphate dehydrogenase. Journal of Applied Biological Sciences, 10(2): 46-50.
29. Kirici, M., Kirici, M., Beydemir, S. & Bulbul, M. (2017a). 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(12): 7074-7082.
30. Kirici, M., Kirici, M., Demir, Y., Beydemir, S. & Atamanalp, M. (2016a). 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
31. Koca, Y.B., Koca, S., Yıldız, S., Gürcü, B., Osanç, E., Tunçbaş, O. & Aksoy, G. (2005). Investigation of histopathological and cytogenetic effects on Lepomis gibbosus (Pisces: Perciformes) in the Çine stream (Aydın/Turkey) with determination of water pollution. Environmental Toxicology, 20(6): 560-571. https://doi.org/10.1002/tox.20145
32. 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. https://doi.org/10.1016/j.etap.2016.04.011
33. Laemmli, D. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227(5259): 680-685. https://doi.org/10.1038/227680a0
34. Ninfali, P., Orsenigo, T., Barociani, L. & Rapa, S. (1990). Rapid purification of glucose-6-phosphate dehydrogenase from mammal's erythrocytes. Preparative Biochemistry & Biotechnology, 20(3-4): 297-309. https://doi.org/ 10.1080/00327489008050202
35. Özer, N., Bilgi, C. & Ögüs, H. (2002). Dog liver glucose-6-phosphate dehydrogenase purification and kinetic properties. The International Journal of Biochemistry and Cell Biology, 34(3): 253–262. https://doi.org/10.1016 /S1357-2725(01)00125-X
36. Patrinostro, X., Carter, M. L., Kramer, A. C. & Lund, T. C. (2013). A Model of glucose-6-phosphate dehydrogenase deficiency in the Zebrafish. Experimental Hematology, 41(8): 697-710. https://doi.org/10.1016/j.exphem.2013. 04.002
37. Rainbow, P. S. & White, S. L. (1990). Comparative accumulating of cobalt by three crustaceans: a decapod, an amphipod and a barnace. Aquatic Toxicology, 16(2): 113-126. https://doi.org/10.1016/0166-445X(90)90081-Y
38. Riganti, C., Gazzano, E., Polimeni, M., Aldieri, E. & Ghigo, D. (2012). The pentose phosphate pathway: an antioxidant defense and a crossroad in tumor cell fate. Free Radical Biology and Medicine, 53(3): 421-436. https://doi.org/10.1016/j.freeradbiomed.2012.05.006
39. Şentürk, M., Ceyhun, S. B., Erdoğan, O. & Küfrevioğlu, Ö. İ. (2009). In vitro and in vivo effects of some pesticides on glucose-6-phosphate dehydrogenase enzyme activity from rainbow trout (Oncorhynchus mykiss) erythrocytes. Pesticide Biochemistry and Physiology, 95(2): 95–99. https://doi.org/10.1016/j.pestbp.2009.07. 005
40. Şentürk, M., Ekinci, D., Alıcı, H. A. & Beydemir, Ş. (2011). Paraoxonase-1, an organophosphate detoxifier and cardio protective enzyme, is inhibited by anesthetics: An in vitro and in vivo insight. Pesticide Biochemistry and Physiology, 101(3): 206-211. https://doi.org/10.1016/j.pestbp.2009.07.005
41. Sivaperumal, T. V., Sankar, P. G. & Nair P. G. V. (2006). Heavy metal concentrations in fish: shellfish and fish products from internal markets of India vis-a-vis international standards. Food Chemistry, 102(3): 612-620. https://doi.org/10.1016/j.foodchem.2006.05.041
42. Söyüt, H., Beydemir, Ş., Ceyhun, S. B., Erdoğan, O. & Kaya, E. D. (2012). Changes in carbonic anhydrase activity and gene expression of Hsp70 in rainbow trout (Oncorhynchus mykiss) muscle after exposure to some metals. Turkish Journal of Veterinary and Animal Science, 36(5): 499-508.
43. Srivastava, S. K. & Beutler, E. (1970). Glutathione metabolism of the erythrocyte. The enzymic cleavage of glutathione–haemoglobin preparations by glutathione reductase. Biochemical Journal, 119(3): 353-357. https://doi.org/10.1042/bj1190353
44. Türkeş, C., Söyüt, H. and Beydemir, S. (2015). Human serum paraoxonase-1 (hPON1): In vitro inhibition effects of moxifloxacin hydrochloride, levofloxacin hemihidrate, cefepime hydrochloride, cefotaxime sodium, and ceftizoxime sodium. Journal of Enzyme Inhibition and Medicinal Chemistry 30(4): 622-628. https://doi.org/10.3109/14756366.2014.959511
45. Türkoğlu, V., Aldemir, S. and Çiftçi, M. (2003). Purification and characterization of glucose 6-phosphate dehydrogenase from sheep liver. Turkish Journal of Chemistry, 27(3): 395–402.
46. Yilmaz, H., Ciftci, M., Beydemir, S. and Bakan, E. (2002). Purification of glucose 6-phosphate dehydrogenase from chicken erythrocytes investigation of some kinetic properties. Preparative Biochemistry and Biotechnology, 32(3), 287–301. https://doi.org/10.1081/PB-120013475
47. Yoshida, A. & Huang, I. Y. (1986). Structure of Human G6PD. London, UK: Academic Press, 86p.