Purification of Glutathione Reductase Enzyme from Whiting Fish Gill Tissue and Investigation of Metal Inhibition
Year 2023,
, 610 - 614, 01.11.2023
Seyit Batuhan Dudak
,
Kübra Işık
,
Betül Mitrovica
,
Ercan Soydan
,
Deniz Ekinci
Abstract
Pollution in the seas basically accumulates in marine organisms. Heavy metal residues in aquatic ecosystems can pass through food and cause toxic effects and accumulations in human health. Glutathione reductase (GR), which is among the basic enzymes, has an important place in the suppression of stress in the cell. In this study, glutathione reductase enzyme from whiting fish gill tissue was partially purified for the first time in the literature and the effects of heavy metal compounds on enzyme activity were determined. The purification process was carried out in three stages as homogenate preparation, ammonium sulfate precipitation and also dialysis. In conclusion the study, optimum level pH 7.0, optimal substrate concentration 2 mM NADPH and optimum buffer 150 mM KH2PO4 were determined. After partial purification, the inhibition effects of Cd2+, Ni2+, Zn2+ as heavy metal ions were investigated. The IC50 levels of heavy metals were calculated as 20.17 µM, 33.7 µM and 59.31 µM, respectively.
References
- Acan NL, Tezcan EF. 1989. Sheep brain glutathione reductase: purification and general properties. FEBS Letters, 250(1): 72-74.
- Açan NL. 1990. Koyun beyni glutatyon redüktazının saflaştırılması ve bazı özelliklerinin incelenmesi. PhD Thesis, Ankara University, Institute of Science, Ankara, Türkiye, pp: 104.
- Angelucci F, Miele AE, Boumis G, Dimastrogiovanni D, Brunori M, Bellelli A. 2008. Glutathione reductase and thioredoxin reductase at the crossroad: the structure of Schistosoma mansoni thioredoxin glutathione reductase. ProtStruct Funct Bioinformat, 72(3): 936-945.
- Blokhina O, Virolainen E, Fagerstedt KV. 2003. Antioxidants, oxidative damage and oxygen deprivation stress: a review. Ann Botany, 91: 179-194.
- Çakmak R, Durdagi S, Ekinci D, Şentürk M, Topal G. 2011. Design, synthesis and biological evaluation of novel nitroaromatic compounds as potent glutathione reductase inhibitors. Bioorg Medic Chem Lett, 21(18): 5398-5402.
- Carlberg I, Mannervik B. 1985. Glutathione reductase. Methods Enzymol, 113: 484-490.
- Çoban TA, Şentürk M, Çiftçi M, Küfrevioğlu OI. 2007. Effects of some metal ions on human erythrocyte glutathione reductase: an in vitro study. Prot Peptide Lett, 14(10): 1027-1030.
- Ekinci D, Ceyhun SB, Şentürk M, Erdem D, Küfrevioğlu Öİ, Supuran CT. 2011. Characterization and anions inhibition studies of an α-carbonic anhydrase from the teleost fish Dicentrarchuslabrax. Bioorg Medic Chem, 19(2): 744-748.
- Erat M. 2002. Purification and characterization of human and bovine erythrocytes glutathione reductase, investigation of inhibition or activation effects of some drugs. PhD Thesis,AtatürkUniversity, Institute of Science, Erzurum, Türkiye, pp: 172.
- Fisher W. 1973. FAO species identification sheets for fishery purposes mediterranean and black sea (Fishing Area 37). FAO, Rome, Italy.
- Işık K, Soydan E. 2023a. Purification and characterisation of glutathione reductase from scorpionfish (scorpaena porcus) and investigation of heavy metal ions inhibition. J Enzyme Inhibit Medic Chem, 38(1): 2167078.
- Işık K, Soydan E. 2023b. Purification and metal inhibition of glutathionereductase enzyme from gill tissue of scorpion fish. Anadolu Tar Bil Derg, 38(1): 221-233.
- Ivanov L. 1985. The fisheries resources of the Mediterranean. Part two: The Black Sea. Stud Rev Gen Fish Council Medit.
- Kocaoğlu E, Talaz O, Çavdar H, Şentürk M, Supuran CT, Ekinci D. 2019. Determination of the inhibitory effects of N-methylpyrrole derivatives on glutathione reductase enzyme. J Enzyme Inhibit Medic Chem, 34(1): 51-54.
- Lue SC. 2009. Regulation of glutathione synthesis. Molec AspectsMedic, 30(1-2): 42-59.
- Macmillan-Crow LA, Cruthirds DL. 2001. Invited review: manganese superoxide dismutase in disease. Free Radical Res, 34(4):325-336.
- Öğüs H, Özer N. 1991. Human jejunal glutathione reductase: purification and evaluation of the NADPH-and glutathione-induced changes in redox state, Biochem MedicMetab Biol, 45(1): 65-73.
- Ogus IH, Ozer N. 1998. Purification of NADPH-free glutathione disulfide reductase from human erythrocytes. Prot Expr Purif, 13(1): 41-44. DOI:10.1006/prep.1997.0865.
- Perry G, Cash AD, Srinivas R, Smith MA. 2002. Metals and oxidative homeostasis in Alzheimer's disease. Drug Devel Res, 56(3): 293-299.
- Sen S, Chakraborty R, Sridhar C, Reddy YSR, De B. 2010. Free radicals, antioxidants, diseases and phytomedicines: Current status and future prospect. Int J Pharmaceut Sci Res. 3(1): 91-100.
- Şentürk E, Şentürk M. 2020. Investigation of some corticosteroids as glutathione reductase inhibitor. IntJ Second Metabol, 7(2): 119-125.
Slastenenko E. 1956. Karadeniz havzası balıkları. E.B.K, İstanbul, Türkiye, pp: 711.
- 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, Comparat Biochem Physiol C-Toxicol Pharmacol, 148(2): 117-121. DOI: 10.1016/j. cbpc.2008.04.005.
- Temel Y, Bengü AŞ, Akkoyun HT, Akkoyun M, Ciftci M. 2017. Effect of astaxanthin and aluminum chloride on erythrocyte G6PD and 6PGD enzyme activities in vivo and on erythrocyte G6PD in vitro in rats. JBiochem Molec Toxicol, 31(10): e21954.
- Toribio F, Martinet LE, Pascual P, Lopez BJ. 1996. Methods for purification of glutathione peroxidase and related enzymes, J Chromatography B, 684: 77-97.
- Tüzen M. 2003. Determination of heavy metals in fish samples of the middle Black Sea (Turkey) by graphite furnace atomic absorption spectrometry. Food Chem, 80(1): 119-123.
- Ulusu G, Erat M, Ciftci M, Sakiroglu H,Bakan E. 2005. Purification and characterization of glutathione reductase from sheep liver. Turkish J VetAnim Sci, 29(5): 1109-1117.
- Valko M, Rhodes CJB, Moncol J, Izakovic MM, Mazur M. 2006. Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chemico-biolInteract, 160(1): 1-40.
- Williams Jr CH. 1976. 3 Flavin-containing dehydrogenases. Enzymes, 13: 89-173.
- Willmore WG, Storey KB. 2007. Purification and properties of glutathione reductase from liver of the anoxiatolerant turtle, Trachemys scripta elegans. Molec Cellular Biochem, 297(1-2): 139-149.
- Wu G, Fang YZ, Yang S, Lupton JR, Turner ND. 2004. Glutathione metabolism and its implications for health. JNutrit, 134(3): 489-492.
Year 2023,
, 610 - 614, 01.11.2023
Seyit Batuhan Dudak
,
Kübra Işık
,
Betül Mitrovica
,
Ercan Soydan
,
Deniz Ekinci
References
- Acan NL, Tezcan EF. 1989. Sheep brain glutathione reductase: purification and general properties. FEBS Letters, 250(1): 72-74.
- Açan NL. 1990. Koyun beyni glutatyon redüktazının saflaştırılması ve bazı özelliklerinin incelenmesi. PhD Thesis, Ankara University, Institute of Science, Ankara, Türkiye, pp: 104.
- Angelucci F, Miele AE, Boumis G, Dimastrogiovanni D, Brunori M, Bellelli A. 2008. Glutathione reductase and thioredoxin reductase at the crossroad: the structure of Schistosoma mansoni thioredoxin glutathione reductase. ProtStruct Funct Bioinformat, 72(3): 936-945.
- Blokhina O, Virolainen E, Fagerstedt KV. 2003. Antioxidants, oxidative damage and oxygen deprivation stress: a review. Ann Botany, 91: 179-194.
- Çakmak R, Durdagi S, Ekinci D, Şentürk M, Topal G. 2011. Design, synthesis and biological evaluation of novel nitroaromatic compounds as potent glutathione reductase inhibitors. Bioorg Medic Chem Lett, 21(18): 5398-5402.
- Carlberg I, Mannervik B. 1985. Glutathione reductase. Methods Enzymol, 113: 484-490.
- Çoban TA, Şentürk M, Çiftçi M, Küfrevioğlu OI. 2007. Effects of some metal ions on human erythrocyte glutathione reductase: an in vitro study. Prot Peptide Lett, 14(10): 1027-1030.
- Ekinci D, Ceyhun SB, Şentürk M, Erdem D, Küfrevioğlu Öİ, Supuran CT. 2011. Characterization and anions inhibition studies of an α-carbonic anhydrase from the teleost fish Dicentrarchuslabrax. Bioorg Medic Chem, 19(2): 744-748.
- Erat M. 2002. Purification and characterization of human and bovine erythrocytes glutathione reductase, investigation of inhibition or activation effects of some drugs. PhD Thesis,AtatürkUniversity, Institute of Science, Erzurum, Türkiye, pp: 172.
- Fisher W. 1973. FAO species identification sheets for fishery purposes mediterranean and black sea (Fishing Area 37). FAO, Rome, Italy.
- Işık K, Soydan E. 2023a. Purification and characterisation of glutathione reductase from scorpionfish (scorpaena porcus) and investigation of heavy metal ions inhibition. J Enzyme Inhibit Medic Chem, 38(1): 2167078.
- Işık K, Soydan E. 2023b. Purification and metal inhibition of glutathionereductase enzyme from gill tissue of scorpion fish. Anadolu Tar Bil Derg, 38(1): 221-233.
- Ivanov L. 1985. The fisheries resources of the Mediterranean. Part two: The Black Sea. Stud Rev Gen Fish Council Medit.
- Kocaoğlu E, Talaz O, Çavdar H, Şentürk M, Supuran CT, Ekinci D. 2019. Determination of the inhibitory effects of N-methylpyrrole derivatives on glutathione reductase enzyme. J Enzyme Inhibit Medic Chem, 34(1): 51-54.
- Lue SC. 2009. Regulation of glutathione synthesis. Molec AspectsMedic, 30(1-2): 42-59.
- Macmillan-Crow LA, Cruthirds DL. 2001. Invited review: manganese superoxide dismutase in disease. Free Radical Res, 34(4):325-336.
- Öğüs H, Özer N. 1991. Human jejunal glutathione reductase: purification and evaluation of the NADPH-and glutathione-induced changes in redox state, Biochem MedicMetab Biol, 45(1): 65-73.
- Ogus IH, Ozer N. 1998. Purification of NADPH-free glutathione disulfide reductase from human erythrocytes. Prot Expr Purif, 13(1): 41-44. DOI:10.1006/prep.1997.0865.
- Perry G, Cash AD, Srinivas R, Smith MA. 2002. Metals and oxidative homeostasis in Alzheimer's disease. Drug Devel Res, 56(3): 293-299.
- Sen S, Chakraborty R, Sridhar C, Reddy YSR, De B. 2010. Free radicals, antioxidants, diseases and phytomedicines: Current status and future prospect. Int J Pharmaceut Sci Res. 3(1): 91-100.
- Şentürk E, Şentürk M. 2020. Investigation of some corticosteroids as glutathione reductase inhibitor. IntJ Second Metabol, 7(2): 119-125.
Slastenenko E. 1956. Karadeniz havzası balıkları. E.B.K, İstanbul, Türkiye, pp: 711.
- 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, Comparat Biochem Physiol C-Toxicol Pharmacol, 148(2): 117-121. DOI: 10.1016/j. cbpc.2008.04.005.
- Temel Y, Bengü AŞ, Akkoyun HT, Akkoyun M, Ciftci M. 2017. Effect of astaxanthin and aluminum chloride on erythrocyte G6PD and 6PGD enzyme activities in vivo and on erythrocyte G6PD in vitro in rats. JBiochem Molec Toxicol, 31(10): e21954.
- Toribio F, Martinet LE, Pascual P, Lopez BJ. 1996. Methods for purification of glutathione peroxidase and related enzymes, J Chromatography B, 684: 77-97.
- Tüzen M. 2003. Determination of heavy metals in fish samples of the middle Black Sea (Turkey) by graphite furnace atomic absorption spectrometry. Food Chem, 80(1): 119-123.
- Ulusu G, Erat M, Ciftci M, Sakiroglu H,Bakan E. 2005. Purification and characterization of glutathione reductase from sheep liver. Turkish J VetAnim Sci, 29(5): 1109-1117.
- Valko M, Rhodes CJB, Moncol J, Izakovic MM, Mazur M. 2006. Free radicals, metals and antioxidants in oxidative stress-induced cancer. Chemico-biolInteract, 160(1): 1-40.
- Williams Jr CH. 1976. 3 Flavin-containing dehydrogenases. Enzymes, 13: 89-173.
- Willmore WG, Storey KB. 2007. Purification and properties of glutathione reductase from liver of the anoxiatolerant turtle, Trachemys scripta elegans. Molec Cellular Biochem, 297(1-2): 139-149.
- Wu G, Fang YZ, Yang S, Lupton JR, Turner ND. 2004. Glutathione metabolism and its implications for health. JNutrit, 134(3): 489-492.