In vitro effects of some metal ions on glutathione reductase in the gills and liver of Capoeta trutta
AbstractMany aquatic environmental problems have arisen in consequence of contamination of water by toxic metals and organic pollutants in the present age of technology. Metals play vital roles in enzyme activities and other metabolic events due to their bioaccumulative and nonbiodegradable properties among aquatic pollutants. The aim of this study was to evaluate the inhibitory effects of some metal ions (Ag+, Cu2+, Co2+, Ni2+, Pb2+ and Zn2+) on Capoeta trutta gill and liver glutathione reductase (EC: 22.214.171.124; GR). For this purpose, initially, GR was purified from C. trutta gill and liver. Purification procedure consisted of three steps; preparation of hemolysate, ammonium sulphate precipitation and 2’, 5’-ADP Sepharose 4B affinity chromatography. Using this procedure, C. turtta gill GR, having the specific activity of 19.111 EU/mg proteins, was purified with a yield of 38.8% and 910.05-fold; C. trutta liver GR, having the specific activity of 16.167 EU/mg proteins, was purified with a yield of 21.1% and 734.86-fold. The purity of the enzymes was checked on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and each purified enzyme showed a single band on the gel. In addition, inhibitory effects of some metal ions (Ag+, Cu2+, Co2+, Ni2+, Pb2+ and Zn2+) on GR from gill and liver were investigated in vitro. Ki constants and IC50 values for metal ions which showed inhibition effects were determined by Lineweaver-Burk graps and plotting activity % vs. [I]. In conclusion, IC50 values for fish gill GR were 0.000625, 0.153, 0.220, 0.247 and 0.216 mM and Ki constants for fish gill GR were 0.00045 ± 0.00008, 0.128 ± 0.036, 0.182 ± 0.138, 0.482 ± 0.219 and 0.112 ± 0.047 mM for Ag+, Cu2+, Co2+, Ni2+, Pb2+ and Zn2+, respectively. IC50 values for fish liver GR were 0.000437, 0.217, 0.185, 0.355 and 0.349 mM and Ki constants for fish liver GR were 0.00025 ± 0.00013, 0.532 ± 0.146, 0.123 ± 0.066, 0.093 ± 0.020 and 0.151 ± 0.084 mM for Ag+, Cu2+, Co2+, Ni2+, Pb2+ and Zn2+, respectively. In vitro inhibition rank order was determined as Ag+ > Co2+ > Zn2+ > Ni2+ > Pb2+ for fish gill GR; Ag+ > Cu2+ > Co2+ > Pb2+ > Ni2+ for fish liver GR. From these results, we showed that Ag+ metal ion is the most potent inhibitor of GR enzyme on gill and liver tissues.
Akerlund, B., Tynell, E., Bratt, G., Bielenstein, M., & Lidman, C. (1997). N-acetyl¬cysteine treatment and the risk of toxic reactions to trimethoprim-sulphamethoxazole in primary Pneumocystis carinii prophylaxis in HIV-infected patients. Journal of Infection, 35, 143–147. >> doi.org/10.1016/S0163-4453(97)91578-4
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, 235–241. >> doi.org/10.3906/sag-1002-4
Akkemik, E., Taser, P., Bayindir, A., Budak, H., & Ciftci, M. (2012). Purification and characterization of glutathione s-transferase from turkey liver and inhibition effects of some metal ions on enzyme activity. Environmental Toxicology and Pharmacology, 34, 888–894. >> doi.org/10.1016/j.etap.2012.08.010
Alim, Z., Camur, B., Beydemir, S., & Kufrevioglu, O. I. (2014). The correlation between some metal concentrations and carbonic anhydrase activity in Tuna (Thunnus thynnus Linnaeus, 1758) gill. Hacettepe Journal of Biology and Chemistry, 42, 219–224.
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, 248–254. >> 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, 5475–5480.
Carlberg, I., & Mannervik, B. (1981). Purification and characterization of glutathi¬one reductase from calf liver. An Improved procedure for affinity chromato¬graphy on 2’,5’-ADP Sepharose 4B. Analytical Biochemistry, 116, 531–536. >> doi.org/10.1016/0003-2697(81)90398-5
Cooper, A. J. L., & Kristal, B. S. (1997). Multiple roles of glutathione in the central nervous system. Biological Chemistry, 378, 793–802.
Ekinci, D., Beydemir, S., & Kufrevioglu, O. I. (2007). In vitro inhibitory effects of some heavy metals on human erythrocyte carbonic anhydrases. Journal of Enzyme Inhibition and Medicinal Chemistry, 22, 745–750. >> doi.org/10.1080/14756360601176048
Farkas, A., Salanki, J., Specziar, A., & Varanka, I. (2001). Metal pollution as health indicator of lake ecosystems. International Journal of Occupational Medicine and Environmental Health, 14, 163–170.
Garcia-Alfonso, C., Martinez-Galisteo, E., Llobell, A., Barcena, J. A., & Lopez-Barea, J. (1993). Horse liver glutathione reductase: Purification and characte¬rization. International Journal of Biochemistry, 25, 61–68. >> doi.org/10.1016/0020-711X(93)90490-6
Gul, M., Kutay, F. Z., Temocin, S., & Hanninen, O. (2000). Cellular and clinical implications of glutathione. Indian Journal of Experimental Biology, 38, 625–634.
Gutterer, J., Dringen, R., Hirrlinger, J., & Hamprect, B. (1999). Purification of glutathione reductase from bovine brain, generation of an antiserum, and immunocytochemical localization of the enzyme in neural cells. Journal of Neurochemistry, 73, 1422–1430. >> doi.org/10.1046/j.1471-4159.1999.0731422.x
Hisar, O., Sonmez, A. Y., Beydemir, S., Hisar, S. A., Yanik, T., & Cronin, T. (2009). Kinetic behaviour of glucose 6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in different tissues of rainbow trout (Oncorhynchus mykiss) exposed to non-lethal concentrations of cadmium. Acta Veterinaria Brno, 78, 179–185. >> doi.org/10.2754/avb200978010179
Isik, M., Demir, Y., Kirici, M., Demir, R., Simsek F., & Beydemir, S. (2015). Changes in the anti-oxidant system in adult epilepsy patients receiving anti-epileptic drugs. Archives of Physiology and Biochemistry, 121, 97–102. >> doi.org/10.3109/13813455.2015.1026912
Jenner, P., & Olanow, C. W. (1998). Understanding cell death in Parkinson’s disease. Annals of Neurology, 44, 72–84. >> doi.org/10.1002/ana.410440712
Kalay, M., & Canli, M. (2000). Elimination of essential (Cu, Zn) and nonessential (Cd, Pb) metals from tissues of a freshwater fish Tilapia zillii following an uptake protocol. Turkish Journal of Zoology, 24, 429–436.
Kalyoncu, L., Kalyoncu, H., & Arslan, G. (2011). Determination of heavy metals and metals levels in five fish species from Işıklı Dam Lake and Karacaören Dam Lake (Turkey). Environmental Monitoring and Assessment, 184, 2231–2235. >> doi.org/10.1007/s10661-011-2112-9
Kaya, E. D., Soyut, H., & Beydemir, S. (2013). Carbonic anhydrase activity from the gilthead seabream (Sparus aurata) liver: The toxicological effects of heavy metals. Environmental Toxicology and Pharmacology, 36, 514–521. >> doi.org/10.1016/j.etap.2013.05.019
Kondo, T., Dale, G. L., & Beutler, E. (1980). Glutathione transport by inside-out vesicles from human erythrocytes. Proceedings of the National Academy of Sciences of the United States of America, 77, 6359–6362.
Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227, 680–683.
Lavery, T. J., Kemper, C. M., Sanderson, K., Schultz, C. G., Coyle, P., Mitchell, J. G., & Seuront, L. (2009). Heavy metal toxicity of kidney and bone tissues in South Australian adult bottlenose dolphins (Tursiops aduncus). Marine Environmental Research, 67, 1–7. >> doi.org/10.1016/j.marenvres.2008.09.005
Le Trang, N., Bhargava, K. K., & Cerami, A. (1983). Purification of glutathione reductase from gerbil liver in two steps. Analytical Biochemistry, 133, 94–99. >> doi.org/10.1016/0003-2697(83)90226-9
Madamanchi, N. R., Anderson, J. V., Alscher, R. G., Cramer, C. L., & Hess, J. L. (1992). Purification of multiple forms of glutathione reductase from pea (Pisum sativum L.) seedlings and enzyme levels in ozone-fumigated pea leaves. Plant Physiology, 100, 138–145. >> doi.org/10.1104/pp.100.1.138
Pacht, E. R., Timerman, A. P., Lykens, M. G., & Merola, A. J. (1991). Deficiency of alveolar fluid glutathione in patients with sepsis and the adult respiratory distress syndrome. Chest, 100, 1397–1403. >> doi.org/10.1378/chest.100.5.1397
Poot, M., Teubert, H., Rabinovitch, P. S., & Kavanagh, T. J. (1995). De novo synthesis of glutathione is required for both entry into and progression through the cell cycle. Journal of Cellular Physiology, 163, 555–560. >> doi.org/10.1002/jcp.1041630316
Raspanti, E., Cacciola, S. O., Gotor, C., Romero, L. C., & Garcia, I. (2009). Impli¬cations of cysteine metabolism in the heavy metal response in Trichoderma harzianum and in three Fusarium species. Chemosphere, 76, 48–54. >> doi.org/10.1016/j.chemosphere.2009.02.030
Sato, M., & Kondoh, M. (2002). Recent studies on metallothionein: Protection against toxicity of heavy metals and oxygen free radicals. Tohoku Journal of Experimental Medicine, 196, 9–22. >> doi.org/10.1620/tjem.196.9
Soyut, H., Beydemir, S., & Hisar, O. (2008). Effects of some metals on carbonic anhydrase from brains of rainbow trout. Biological Trace Element Research, 123, 179–190. >> doi.org/10.1007/s12011-008-8108-9
Squadrone, S., Prearo, M., Brizio, P., Gavinelli, S., Pellegrino, M., Scanzio, T., Guarise, S., Benedetto, A., & Abete, M. C. (2013). Heavy metals distribution in muscle, liver, kidney and gill of european catfish (Silurus glanis) from Italian Rivers. Chemosphere, 90, 358–365. >> doi.org/10.1016/j.chemosphere.2012.07.028
Taser, P., & Ciftci, M. (2012). Purification and characterization of glutathione reductase from turkey liver. Turkish Journal of Veterinary and Animal Sciences, 36, 546–553. >> doi.org/10.3906/vet-1103-5
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, 148, 117–121. >> doi.org/10.1016/j.cbpc.2008.04.005
Van den Dobbelsteen, D. J., Nobel, C. S. I., Schlegel, J., Cotgreave, I. A., Orrenius, S., & Slater, A. F. (1996). Rapid and specific efflux of reduced glutathione during apoptosis induced by Anti-Fas/APO-1 antibody. Journal of Biological Chemistry, 271, 15420–15427. >> doi.org/10.1074/jbc.271.26.15420
Willmore, W. G., & Storey, K. B. (2007). Purification and properties of glutathione reductase from liver of the anoxia-tolerant turtle, Trachemys scripta elegans. Molecular and Cellular Biochemistry, 297, 139–149. >> doi.org/10.1007/s11010-006-9339-8
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, 193–198. >> doi.org/10.1016/j.parint.2012.12.008
Yi, Y. J., & Zhang, S. H. (2012). Heavy metal (Cd, Cr, Cu, Hg, Pb, Zn) concentrations in seven fish species in relation to fish size and location along the Yangtze River. Environmental Science and Pollution Research, 19, 3989–3996. >> doi.org/10.1007/s11356-012-0840-1
Yoshida, K., Hirokawa, J., Tagami, S., Kawakami, Y., Urata, Y., & Kondo, T. (1995). Weakened cellular scavenging activity against oxidative stress in diabetes mellitus: Regulation of glutathione synthesis and efflux. Diabetologia, 38, 201–210. >> doi.org/10.1007/BF00400095
Yousafzai, A. M., Siraj, M., Ahmad, H., & Chivers, D. P. (2012). Bio¬accumulation of heavy metals in common carp: Implications for human health. Pakistan Journal of Zoology, 44, 489–494.
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