The impacts of some metals on the activity of corb gill Umbrina cirrosa Carbonic Anhydrase

Yıl 2014, Cilt: 42 Sayı: 4, 499 - 504, 01.11.2014

Namık Kılınç , Zuhal Alım Mehmet Mustafa İşgör Şükrü Beydemir

Öz

Metal toxicity causes oxidative stress in fish. This situation is a potential risk factor for humans and other living feeding on contaminated fish. In this study, the inhibition effects of heavy metals on carbonic anhy- drase enzyme from the corb fish gill were investigated. The carbonic anhydrase enzyme was purified from gill of corb fish with a specific activity of 2093,9 EUmg−1 and 86,51% yield and approximately 160 fold using Sep- harose 4B–L-tyrosine sulfanilamide affinity chromatography method. SDS–polyacrylamide gel electrophoresis showed a single band corresponding to a molecular weight of approximately 30,8 kDa. Inhibitory effects of metals Ag+, Cu2+, Pb2+, Zn2+, Cd2+, Ni2+ on CA activity were determined at different concentrations using the hydratase method under in vitro conditions. Consequently, in vitro inhibition rank order was determined as Ag+> Cu2+> Pb2+> Zn2+ > Cd2+> Ni2+. From these results, we showed that Ag+ is the most potent inhibitor of CA enzyme.

Anahtar Kelimeler

Carbonic anhydrase, Gill, Heavy metals, Inhibition

Bazı Metallerin Minekop Umbrina Cirrosa Solungacı Karbonik Anhidraz Aktivitesi Üzerine Etkileri

Öz

S Metal toksisitesi balıklarda oksidatif strese sebep olur. Bu durum insanlar ve diğer balık tüketen canlılar için potansiyel bir risk faktörüdür. Bu çalışmada minekop balığı solungacından saflaştırılan karbonik anhidraz enzimi üzerine bazı ağır metallerin inhibisyon etkileri incelendi. Minekop balığı solungacından, karbonik anhidraz, Sepharose 4B–L-tyrosin sulfanilamit afinite kolonu kullanılarak 2093,9 EUmg−1 spesifik aktiviteyle ve 86,51% verimle yaklaşık 160 kat saflaştırıldı. SDS-poliakrilamid jel elektroforezinde tek band gözlendi ve buradan enzimin molekül kütlesi yaklaşık olarak 30,8 kDa olarak belirlendi. Bazı ağır metallerin Ag+, Cu2+, Pb2+, Zn2+, Cd2+, Ni2+ CA enzimi aktivitesi üzerine etkileri hidrataz metodu kullanılarak in vitro olarak incelendi. Sonuç olarak, ağır metallerin inhibisyon etkileri Ag+> Cu2+> Pb2+> Zn2+ > Cd2+> Ni2+ şeklinde belirlendi. Bu sonuçlar Ag+’nın en önemli potansiyel CA inhibitörü olduğunu gösterdi

Anahtar Kelimeler

Karbonik anhidraz, solungaç, ağır metaller, inhibisyon

Kaynakça

• 1. A. Farkas, J. Salanki, A. Specziar, Relation between growth and the heavy metal concentration in organs of bream Abramis brama L. populating lake Balaton. Arch. Environ. Contam. Toxicol., 43 (2002) 236.
• 2. M.H.A. Yousuf, El-Shahawi. Trace metals in Lethrinus lentjan fish from Arabian Gulf: Metal accumulation in Kidney and Heart Tissues. Bull. Environ. Contam. Toxicol., 62 (1999) 293.
• 3. H. Soyut, S. Beydemir, The impact of heavy metals on the activity of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) kidney. Toxicol Ind Health 2011 DOI:10.1177/0748233711410914.
• 4. B.P. Cid, C. Boia, L. Pombo, E. Rebelo, Determination of trace metals in fish species of the Ria de Aveiro (Portugal) by electrothermal atomic absorption spectrometry. Food Chemistry. 75 (2001) 93.
• 5. D. Velez, R. Montoro, Arsenic speciation in manufactured seafood products: a review. J. food. Protect. 61 (1998) 1240.
• 6. H.B. Conacher, B.D. Page, J.J. Ryan, Industrial chemical contamination of foods [Review]. Food Addit. Contam. 10(1993) 129.
• 7. E.O. Farombi, O.A. Adelowo, Y.R. Ajimoko, Biomarkers of oxidative stress and heavy metal levels as indicators of environmental pollution in African Cat fish (Clarias gariepinus) from Nigeria ogun river. Int. J. Environ. Res. Public Health., 4 (2007) 158.
• 8. M.Z. Vosyliene, A. Jankaite, Effect of heavy metal model mixture on rainbow trout biological parameters. Ekologija. 4 (2006) 12.
• 9. W. Ashraj, Accumulation of heavy metals in kidney and heart tissues of Epinephelus microdon fish from the Arabian Gulf. Environ. Monit. Assess. 101 (2005) 311.
• 10. E. Somer, Toxic potential of trace metals in foods. A review. Journal of Food Science. 39 (1974) 215.
• 11. K. Güven, C. Özbay, E. Ünlü, A. Satar, Acute lethal toxicity and accumulation of copper in Gammarus pulex (L.) (Amphipoda). Turkish Journal of Biology. 23 (1999) 510.
• 12. D.J. Jefferies, P. Freestone, Chemical analysis of some coarse fish from a Suffolk River carried out as part of the preparation for the first release of captive-bred otters. J. Otter Trust, 1(8) (1984) 17.
• 13. C. Hogstrand, E.A. Ferguson, F. Galvez, R. Shaw, N.A. Webb, C.M. Wood, Physiology of acute silver toxicity in the starry flounder (Platichthys stellatus) in seawater. J. Comp. Physiol. B. 169 (1999) 461.
• 14. P.S. Basha, A.U. Rani, Cadmium-induced antioxidant defense mechanism in freshwater teleost Oreochromis mossambicus (Tilapia). Ecotoxicol. Environ. Saf. 56 (2003) 218.
• 15. A.A.R. Radi, B. Matkovics, Effects of metal ions on the antioxidant enzyme activities, protein contents and lipid peroxidation of carp tissues. Comp. Biochem. Physiol. C 90 (1988) 69.
• 16. H., Roche, G.Boge, Effects of Cu, Zn and Cr salts on antioxidant enzyme activities in vitro of red blood cells of a marine fish Dicentrarchus labrax. Toxicol. in Vitro. 7 (1993) 623.
• 17. M., Canli, R.M. Stagg, The effects of in vivo exposure to cadmium, copper, and zinc on the activities of gill ATPases in the Norway lobster Nephrops norvegicus. Arch. Environ. Contam. Toxicol. 31 (1996) 491.
• 18. C.K.C. Wong, M.H. Wong, Morphological and biochemical changes in the gills of tilapia (Oreochromis mossambicus) to ambient cadmium exposure. Aquat. Toxicol. 48 (2000) 517.
• 19. M.G.L. Ribeiro, A.R. Pedrenho, Hasson-Voloch, A. Electrocyte (Na+–K+)-ATPase inhibition induced by zinc is reverted by dithiothreitol. The Int. J. Biochem. Cell Biol. 34 (2002) 516.
• 20. C. Dautremepuits, S. Paris-Palacios, S. Betoulle, G. Vernet, Modulation in hepatic and head kidney parameters of carp (Cyprinus carpio L.) induced by copper and chitosan. Comp. Biochem. Physiol. C, 137 (2004) 325.
• 21. K.M. Gilmour, S.F. Perry, Carbonic anhydrase and acid– base regulation in fish. The Journal of Experimental Biology 212 (2009) 1647.
• 22. F.M. El-Demerdash, E.I. Elagamy, Biological effects in Tilapia nilotica fish as indicators of pollution by cadmium and mercury. Int. J. Environ. Health Res. 9 (1999) 173.
• 23. Ş. Gül, E. Belge-Kurutaş, E. Yıldız, A. Şahan, F. Doran, Pollution correlated modifications of liver antioxidant systems and histopathology of fish (Cyprinidae) living in Seyhan Dam Lake, Turkey. Environ. Int. 30 (2004) 605.
• 24. D. Ekinci, S. Beydemir, Z. Alim, Some drugs inhibit in vitro hydratase and esterase activities of human carbonic anhydrase-I and II. Pharmacological Reports 59 (2007a) 580.
• 25. D. Ekinci, S. Beydemir, O.I. Kufrevioglu, In vitro inhibitory effects of some heavy metals on human erythrocyte carbonic anhydrases. Journal of Enzyme Inhibition and Medicinal Chemistry 22 (2007b) 745.
• 26. D. Ekinci, S. Beydemir, Risk assessment of pesticides and fungicides for acid–base regulation and salt transport in rainbow trout tissues. Pesticide Biochemistry and Physiology. 97 (2010) 66.
• 27. H. Soyut, S. Beydemir, O. Hisar, Effects of some metals on carbonic anhydrase from brains of rainbow trout. Biol. Trace Elem. Res., 123 (2008) 179.
• 28. H. Soyut, S. Beydemir, Purification and some kinetic properties of carbonic anhydrase from rainbow trout (Oncorhynchus mykiss) liver and metal inhibition. Protein Peptide Lett. 15 (2008) 528.
• 29. E.D. Kaya, H. Söyüt, S. Beydemir, Carbonic anhydrase activity from the gilthead sea bream (Sparus aurata) liver: the toxicological effects of heavy metals. Environ Toxicol Pharmacol. 36(2) (2013) 514.
• 30. K.M Wilbur, N.G. Anderson, Electrometric and colorimetric determination of carbonic anhydrase. J. Biol. Chem. 176 (1948) 147.
• 31. M.M. Bradford, A rapid and sensitive method for the quantition of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72 (1976) 248.
• 32. D.K. Laemmli, Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227 (1970) 680.
• 33. S. Sender, K. Böttcher, Y. Cetin, G. Gros, Carbonic Anhydrase in the Gills of Seawaterand Freshwateracclimated Flounders Platichthys flesus: Purification, Characterization, and Immunohistochemical Localization. The Journal of Histochemistry & Cytochemistry 47 (1999) 43.
• 34. S.F. Perry, P. Laurent, The role of carbonic anhydrase in carbon dioxide excretion, acid-base balance and ionic regulation in aquatic gill breathers. In Truchot J-P, Lahlou B, eds. Animal Nutrition and Transport Processes. 2. Transport, Respiration and Excretion: Comparative and Environmental Aspects. 6 (1990) 39.