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Partial Purification and Some Interesting Properties of Glutathione Peroxidase from Liver of Camel (Camelus dromedarius)

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Abstract

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 β-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.

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Abbreviations

ROS:

reactive oxygen species

GPx:

glutathione peroxidase

GSH:

reduced glutathione

DTT:

D,L-dithiothreitol

PMSF:

phenylmethylsulfonylfluoride

DTNB:

5,5′-dithiobis(2-nitrobenzoic acid)

NBT:

nitro blue tetrazolium

PMS:

phenazinemethosulphate

CHP:

cumenehydroperoxide

TBH:

tert-butyl hydroperoxide

BP:

benzoyl peroxide

Cys:

L-cysteine

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

EDTA:

ethylenediaminetetraacetic acid

β-ME:

β-mercaptoethanol

ICP-MS:

inductively coupled plasma mass spectrometry

TCA:

trichloroacetic acid

Se-GPx:

selenium-dependent GPx

non-Se-GPx:

non-selenium-dependent GPx

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Authors and Affiliations

  1. Laboratory of Biochemistry and Neuroscience, Team of Applied Biochemistry and Toxicology, Faculty of Science and Technology, University Hassan First, Settat, 26000, Morocco

    Abdelbasset Chafik & Abdelkhalid Essamadi

  2. College of Education, Bayburt University, Bayburt, 69000, Turkey

    Safinur Yildirim Çelik

  3. Department of Nanoscience and Nanoengineering, Institute of Science, Atatürk University, Erzurum, 25240, Turkey

    Kübra Solak

  4. Chemistry Education, Kazim Karabekir Education Faculty, Atatürk University, Erzurum, 25240, Turkey

    Ahmet Mavi

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  1. Abdelbasset Chafik
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  2. Abdelkhalid Essamadi
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Chafik, A., Essamadi, A., Çelik, S.Y. et al. Partial Purification and Some Interesting Properties of Glutathione Peroxidase from Liver of Camel (Camelus dromedarius). Russ J Bioorg Chem 44, 41–51 (2018). https://doi.org/10.1134/S1068162018010041

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