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- Title
Purification and biochemical characterization of a secreted group IIA chicken intestinal phospholipase A<sub>2</sub>.
- Authors
Karray, Aida; Frikha, Fakher; Ali, Yassine Ben; Gargouri, Youssef; Bezzine, Sofiane
- Abstract
Background: Secretory phospholipase A2 group IIA (IIA PLA2) is a protein shown to be highly expressed in the intestine of mammals. However, no study was reported in birds. Results: Chicken intestinal group IIA phospholipase A2 (ChPLA2-IIA) was obtained after an acidic treatment (pH.3.0), precipitation by ammonium sulphate, followed by sequential column chromatographies on Sephadex G-50 and mono-S ion exchanger. The enzyme was found to be a monomeric protein with a molecular mass of around 14 kDa. The purified enzyme showed a substrate preference for phosphatidylethanolamine and phosphatidylglycerol, and didn't hydrolyse phosphatidylcholine. Under optimal assay conditions, in the presence of 10 mM NaTDC and 10 mM CaCl2, a specific activity of 160 U.mg-1 for purified ChPLA2-IIA was measured using egg yolk as substrate. The fifteen NH2-terminal amino acid residues of ChPLA2-IIA were sequenced and showed a close homology with known intestinal secreted phospholipases A2. The gene encoding the mature ChPLA2-IIA was cloned and sequenced. To further investigate structure-activity relationship, a 3D model of ChPLA2-IIA was built using the human intestinal phospholipase A2 structure as template. Conclusion: ChPLA2-IIA was purified to homogeneity using only two chromatographic colomns. Sequence analysis of the cloned cDNA indicates that the enzyme is highly basic with a pI of 9.0 and has a high degree of homology with mammalian intestinal PLA2-IIA.
- Subjects
PHOSPHOLIPASE A2; PROTEINS; MAMMALS; AMMONIUM sulfate; AMINO acids
- Publication
Lipids in Health & Disease, 2011, Vol 10, Issue 1, p27
- ISSN
1476-511X
- Publication type
Article
- DOI
10.1186/1476-511X-10-27