Yazar "Çelik S.Y." seçeneğine göre listele
Listeleniyor 1 - 4 / 4
Sayfa Başına Sonuç
Sıralama seçenekleri
Öğe Characterization of an interesting selenium-dependent glutathione peroxidase (Se-GPx) protecting cells against environmental stress: The Camelus dromedarius erythrocytes Se-GPx(Elsevier Ltd, 2019) Chafik A.; Essamadi A.; Çelik S.Y.; Solak K.; Mavi A.Camel lives most of its life under high environmental stress in the desert. Glutathione peroxidase plays a key role in protecting cells against oxidative stress. For the first time, selenium-dependent glutathione peroxidase (Se-GPx) was purified from camel erythrocytes, biochemically characterized, and some of its properties were studied. The enzyme was purified using ethanol-chloroform treatment, acetone precipitation and ion exchange chromatography. A purification fold of 33.72 with 0.19% yield were obtained. The native molecular weight of the enzyme was estimated to be about 69 kDa. On SDS-PAGE, the enzyme was composed of two different subunits with a molecular weight of approximately 53 and 21 kDa. An optimum temperature of 47 °C and an optimum pH of 7.2 were found. The activation energy was 41.71 kJ/mol. This enzyme was inhibited by thiol reagents, D,L-Dithiothreitol and ?-Mercaptoethanol, and was sensitive to bivalent cations. The enzyme had a general specificity toward hydroperoxides, and high specificity for reduced glutathione. The purified enzyme contained 3.06 mol of selenium per mol of protein. The Km and Vmax values for hydrogen peroxide and reduced glutathione were 0.72 and 1.58 mM, and 25.33 and 31.03 U/mg, respectively. The biochemical properties of camel Se-GPx were different comparing to those of mammalian species. Lower molecular weight, heterodimeric structure, higher optimum temperature, relatively lower optimum pH, lower content of selenium and higher affinity for hydrogen peroxide at low reduced glutathione concentration, these could be explained by the fact that camel is able to live in the intense environmental stress in the desert. © 2019 Elsevier LtdÖğe Partial Purification and Some Interesting Properties of Glutathione Peroxidase from Liver of Camel (Camelus dromedarius)(Pleiades Publishing, 2018) Chafik A.; Essamadi A.; Çelik S.Y.; Solak K.; Mavi A.Climate change and increasing temperatures are global concerns. Well adapted to desert life, the camel (Camelus dromedarius) lives most of its life under high environmental stress and represents an ideal model for studying desert adaptation among mammals. Glutathione peroxidase is the principal antioxidant defense system capable of protecting cells from oxidative stress. Glutathione Peroxidase from camel liver was purified (11.64-fold purification with 1.73% yield) and characterized The molecular weight of the enzyme was estimated to be about 69 kDa by gel filtration and 34 kDa by SDS-PAGE, implying dimeric structure of the protein. An optimum temperature of 47°C and an optimum pH of 7.8 were found. This enzyme is a typical SH-enzyme that is inhibited by D,L-dithiothreitol and ?-mercaptoethanol and sensitive to bivalent cations. The enzyme had common specificity toward hydroperoxides and high specificity for reduced glutathione. The Km and Vmax values for hydrogen peroxide and reduced glutathione were 0.57 and 2.10 mM and 1.11 and 0.87 U/mg, respectively. The purified enzyme contained 16 ng of selenium per mg of protein. Our results show that the camel glutathione peroxidse exhibits properties different of those reported for other mammalian species. Lower molecular weight, homodimeric structure, higher optimum temperature, relatively low optimum pH, high affinity for hydrogen peroxide at low concentration of reduced glutathione and very low content of selenium could be explained by adaptation of the camel to living in the desert under intense environmental stress. © 2018, Pleiades Publishing, Ltd.Öğe Purification and biochemical characterization of a novel copper, zinc superoxide dismutase from liver of camel (Camelus dromedarius): An antioxidant enzyme with unique properties(Academic Press Inc., 2019) Chafik A.; Essamadi A.; Çelik S.Y.; Mavi A.A novel copper, zinc superoxide dismutase (CuZnSOD) was purified to homogeneity from the liver of an animal well adapted to the stressful living conditions of the desert, the camel (Camelus dromedarius). The biochemical properties of camel liver CuZnSOD were examined. The purified enzyme had a native molecular weight of 28 kDa, as judged by gel filtration chromatography, and showed a single band at 27 kDa on SDS-PAGE, indicating that it is a monomeric protein. Optimal activity of the purified enzyme occurred at 43 °C and pH 6.0, and the activation energy was 1.42 kJ/mol. CuZnSOD activity was strongly inhibited by ?-ME, DTT, H 2 O 2 and SDS and slightly inhibited by EDTA, NaN 3 and PMSF. Al 3+ , Ca 2+ , Cd 2+ , Mg 2+ and Zn 2+ stimulated CuZnSOD activity, whereas Ba 2+ , Co 2+ , Fe 2+ and Ni 2+ inhibited it. The purified enzyme contained 0.010 µg of Cu and 0.69 µg of Zn per mg of protein. K m , V max , k cat and k cat /K m values for NBT and riboflavin were 16.27 and 0.16 µM, 20.85 and 21.54 U/mg, 9.65 and 9.97 s ?1 , and 0.59 and 62.33 s -1 µM ?1 , respectively. Camel liver CuZnSOD exhibited unique biochemical properties compared to those of other CuZnSODs, including lower molecular weight with a monomeric structure, higher optimum temperature, very low E a , very low optimum pH, very low contents of Cu and Zn, and higher affinity, turnover number and catalytic efficiency for riboflavin. These unique properties of camel liver CuZnSOD might be related to the ability of this animal to inhabit stressful desert conditions. © 2019 Elsevier Inc.Öğe Purification of camel liver catalase by zinc chelate affinity chromatography and pH gradient elution: An enzyme with interesting properties(Elsevier B.V., 2017) Chafik A.; Essamadi A.; Çelik S.Y.; Mavi A.Climate change and increasing temperatures are global concerns. Camel (Camelus dromedarius) lives most of its life under high environmental stress in the desert and represent ideal model for studying desert adaptation among mammals. Catalase plays a key role in protecting cells against oxidative stress. For the first time, catalase from camel liver was purified to homogeneity by zinc chelate affinity chromatography using pH gradient elution, a better separation was obtained. A purification fold of 201.81 with 1.17% yield and a high specific activity of 1132539.37 U/mg were obtained. The native enzyme had a molecular weight of 268 kDa and was composed of four subunits of equal size (65 kDa). The enzyme showed optimal activity at a temperature of 45 °C and pH 7.2. Thiol reagents, ?-Mercaptoethanol and D,L-Dithiothreitol, inhibited the enzyme activity. The enzyme was inhibited by Al3+, Cd2+ and Mg2+, whereas Ca2+, Co2+ and Ni2+ stimulated the catalase activity. Reduced glutathione has no effect on catalase activity. The Km and Vmax of the enzyme for hydrogen peroxide were 37.31 mM and 6185157 U/mg, respectively. Sodium azide inhibited the enzyme noncompetitively with Ki value of 14.43 ?M, the IC50 was found to be 16.71 ?M. The properties of camel catalase were different comparing to those of mammalian species. Relatively higher molecular weight, higher optimum temperature, protection of reduced glutathione from hydrogen peroxide oxidation and higher affinity for hydrogen peroxide and sodium azide, these could be explained by the fact that camel is able to live in the intense environmental stress in the desert. © 2017 Elsevier B.V.
