title: Combined proteomic and molecular approaches for cloning and characterization of copper-zinc superoxide dismutase (Cu, Zn-SOD2) from garlic (Allium sativum).
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.
authors.affiliation: Department of Bioengineering, National Institute of Applied Sciences and Technology, University of Carthage, 676-1080 Tunis Cedex, Tunisia. hadjiimen@yahoo.fr
authors.forename: Imen
authors.initials: I
authors.lastname: Hadji Sfaxi
authors.forename: Aymen
authors.initials: A
authors.lastname: Ezzine
authors.forename: Laurent
authors.initials: L
authors.lastname: Coquet
authors.forename: Pascal
authors.initials: P
authors.lastname: Cosette
authors.forename: Thierry
authors.initials: T
authors.lastname: Jouenne
authors.forename: M Nejib
authors.initials: MN
authors.lastname: Marzouki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002457:Cell Extracts; D002853:Chromatography, Liquid; D003001:Cloning, Molecular / Q000379:methods*; D019610:Cytoprotection; D004317:Doxorubicin / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D015180:Electrophoresis, Gel, Two-Dimensional; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005737:Garlic / Q000201:enzymology* / Q000235:genetics*; D013058:Mass Spectrometry; D008546:Melanoma, Experimental / Q000473:pathology; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D016133:Polymerase Chain Reaction; D020033:Protein Isoforms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013482:Superoxide Dismutase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D034421:Sus scrofa
chemical_list: D002457:Cell Extracts; D010455:Peptides; D020033:Protein Isoforms; D004317:Doxorubicin; D013482:Superoxide Dismutase