Purification and biochemical characterization of phospholipase A2 from dromedary pancreas

被引:18
作者
Ben Bacha, Abir
Gargouri, Youssef
Bezzine, Sofiane
Mejdoub, Hafedh
机构
[1] ENIS, Lab Biochim & Genie Enzymat Lipases, Sfax 3038, Tunisia
[2] Fac Sci Sfax, UCSR Sequenceure Prot, Sfax 3038, Tunisia
来源
BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS | 2006年 / 1760卷 / 08期
关键词
pancreatic phospholipase; PC; bile salts; Ca2+; sequence;
D O I
10.1016/j.bbagen.2006.03.014
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Dromedary pancreatic PLA2 (DrPLA2) was purified from delipidated pancreases. Pure protein was obtained after heat and acidic treatment (70 degrees 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 degrees 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. (c) 2006 Elsevier B.V. All rights reserved.
引用
收藏
页码:1202 / 1209
页数:8
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