Purification and biochemical characterization of phospholipase A2 from dromedary pancreas

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• 作者：Abir Ben Bacha ; Youssef Gargouri ; Sofiane Bezzine and Hafedh Mejdoub
• 关键词：Pancreatic phospholipase ; PC ; Bile salts ; Ca²⁺ ; Sequence
• 刊名：Biochimica et Biophysica Acta (BBA)/General Subjects
• 出版年：2006
• 出版时间：August 2006
• 年：2006
• 卷：1760
• 期：8
• 页码：1202-1209
• 全文大小：351 K

文摘

Dromedary pancreatic PLA2 (DrPLA2) was purified from delipidated pancreases. Pure protein was obtained after heat and acidic treatment (70 °C; pH 3.0), precipitation by ammonium sulphate and ethanol respectively, followed by sequential column chromatographies on Sephadex G-50, MonoS Sepharose, MonoQ Sepharose and C-8 reverse phase high pressure liquid chromatography. Purified DrPLA2, which is not glycosylated protein, was found to be monomeric protein with a molecular mass of 13748.55 Da. A specific activity of 600 U/mg for purified DrPLA2 was measured at optimal conditions (pH 8.0 and 37 °C) in the presence of 3 mM NaTDC and 7 mM CaCl₂ using PC as substrate. The sequence of the first fourteen amino-acid residues at the N-terminal extremity of DrPLA2 was determined by automatic Edman degradation. One single sequence was obtained and shows a close similarity with all other known pancreatic secreted phospholipases A2.