Two purified and characterized phospholipases A2 from Cerastes cerastes

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• 作者：Raoudha Zouari-Kessentini ; José ; Luis ; Aida Karray ; Olfa Kallech-Ziri ; Najet Srairi-Abid ; Amine Bazaa ; Erwann Loret ; Sofiane Bezzine ; Mohamed El Ayeb ; Naziha Marrakchi
• 关键词：Venom ; Secreted phospholipase A2 ; Cell adhesion ; Cell migration
• 刊名：Toxicon
• 出版年：2009
• 出版时间：15 March 2009
• 年：2009
• 卷：53
• 期：4
• 页码：444-453
• 全文大小：579 K

文摘

Two non-toxic PLA2s were purified to homogeneity from Cerastes cerastes Tunisian snake venom. The purification process employed gel filtration on Sephadex G-75 followed by C18 reverse phase high-pressure liquid chromatography. These two acidic enzymes, namely CC-PLA2-1 and CC-PLA2-2, have a molecular weight of 13,737.52 and 13,705.63 Da, respectively. These two PLA2 are the first reported glycosylated phospholipases A2 purified from snake venom. The rates of glycosylation are 2.5 % and 0.5 % (w/w), respectively. Specific activities of 1800 U/mg and 2400 U/mg for CC-PLA2-1 and CC-PLA2-2, respectively, were measured at optimal conditions. CC-PLA2-1 and CC-PLA2-2 strongly inhibited coagulation. They also exhibited a marked dose-dependent inhibitory effect on platelet aggregation induced by ADP and arachidonic acid in platelet-rich plasma. Interestingly, CC-PLA2-1 and CC-PLA2-2 inhibited in a dose-dependent manner adhesion of IGR39 melanoma and HT1080 fibrosarcoma cells to fibrinogen and fibronectin. Furthermore, both CC-PLA2-1 and CC-PLA2-2 abolished HT1080 cell migration towards fibrinogen and fibronectin. This activity is reported for the first time for PLA2 enzymes.