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Combined Proteomic and Molecular Approaches for Cloning and Characterization of Copper–Zinc Superoxide dismutase (Cu, Zn-SOD2) from Garlic (Allium sativum)

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Abstract

Superoxide dismutases (SODs; EC 1.15.1.1) are key enzymes in the cells protection against oxidant agents. Thus, SODs play a major role in the protection of aerobic organisms against oxygen-mediated damages. Three SOD isoforms were previously identified by zymogram staining from Allium sativum bulbs. The purified Cu, Zn-SOD2 shows an antagonist effect to an anticancer drug and alleviate cytotoxicity inside tumor cells lines B16F0 (mouse melanoma cells) and PAE (porcine aortic endothelial cells). To extend the characterization of Allium SODs and their corresponding genes, a proteomic approach was applied involving two-dimensional gel electrophoresis and LC–MS/MS analyses. From peptide sequence data obtained by mass spectrometry and sequences homologies, primers were defined and a cDNA fragment of 456 bp was amplified by RT-PCR. The cDNA nucleotide sequence analysis revealed an open reading frame coding for 152 residues. The deduced amino acid sequence showed high identity (82–87%) with sequences of Cu, Zn-SODs from other plant species. Molecular analysis was achieved by a protein 3D structural model.

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Acknowledgments

This study was sustained by a financial support of the Bioengineering Laboratory 99UR09-26 (INSAT) from the Tunisian Ministry of High Education Scientific Research.

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Authors and Affiliations

  1. Bioengineering Unit 99UR09-26, Department of Bioengineering, National Institute of Applied Sciences and Technology, University of Carthage, 676-1080, Tunis Cedex, Tunisia

    Imen Hadji Sfaxi, Aymen Ezzine & M. Nejib Marzouki

  2. Laboratory of Polymers, Biopolymers, Surfaces, UMR 6270 CNRS, Proteomic Platform of the IFRMP23, University of Rouen, Rouen, France

    Laurent Coquet, Pascal Cosette & Thierry Jouenne

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  1. Imen Hadji Sfaxi
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  2. Aymen Ezzine
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  3. Laurent Coquet
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Correspondence to Imen Hadji Sfaxi.

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Hadji Sfaxi, I., Ezzine, A., Coquet, L. et al. Combined Proteomic and Molecular Approaches for Cloning and Characterization of Copper–Zinc Superoxide dismutase (Cu, Zn-SOD2) from Garlic (Allium sativum). Mol Biotechnol 52, 49–58 (2012). https://doi.org/10.1007/s12033-011-9473-8

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