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Biochemical and molecular characterization of purified chicken pancreatic phospholipase A 2
Author(s) -
Karray Aida,
Frikha Fakher,
Ben Bacha Abir,
Ben Ali Yassine,
Gargouri Youssef,
Bezzine Sofiane
Publication year - 2009
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2009.07160.x
Subject(s) - phosphatidylcholine , phospholipase , molecular mass , biochemistry , homology (biology) , peptide sequence , chemistry , amino acid , ammonium sulfate precipitation , phospholipase a , biology , microbiology and biotechnology , size exclusion chromatography , phospholipid , chromatography , enzyme , gene , phospholipase a2 , membrane
Chicken pancreatic phospholipase A 2 (ChPLA 2 ) was purified from delipidated pancreases using ammonium sulfate and ethanol precipitation, followed by sequential column chromatography steps on MonoQ Sepharose and size exclusion HPLC columns. ChPLA 2 was found to be a nonglycosylated monomeric protein with a molecular mass of 14 kDa and a specific activity of 400 U·mg −1 in the presence of 1 m m sodium taurodeoxycholate and 4 m m CaCl 2 with phosphatidylcholine as substrate. The N‐terminal sequence of the first 15 amino acids of ChPLA 2 was determined, and showed a high degree of homology with known mammal pancreatic phospholipases A 2 . The gene encoding the mature ChPLA 2 was cloned and sequenced. The deduced amino acid sequence of the mature ChPLA 2 confirmed the high level of identity with mammal pancreatic PLA 2 . To investigate the structure–activity relationships, a 3D model of group IB ChPLA 2 was built using the porcine pancreatic phospholipase A 2 structure as template.