Isolation and characterization of phospholipase A 2, from Agkistrodon bilineatus (common cantil) venom

Abstract

1. 1. Phospholipase A 2 was isolated from Agkistrodon bilineatus venom by Sephadex G-75 and CM-Cellulose column chromatographies. 2. 2. The purified phospholipase A 2-I gave a single band on disc polyacrylamide gel electrophoresis, isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis. 3. 3. The enzyme preparation had a molecular weight of 14,000, isoelectric point of pH 8.77 and possessed 123 amino acid residues. 4. 4. The purified phospholipase A 2 possessed lethal, indirect hemolytic and anticoagulant activities. 5. 5. The enzyme hydrolyzed the phospholipids phosphatidyl choline (PC), phosphatidyl ethanolamine (PE), phosphatidyl inositol (PI) and phosphatidyl serine (PS). 6. 6. The concentration of mouse diaphragm was inhibited and the contraction of guinea pig left atrium was increased by phospholipase A 2-I. 7. 7. Phospholipase A 2 activity of this preparation was inhibited by ethylenediamine tetraacetic acid, p-bromo phenacyl bromide, n-bromo succinimide or dithiothreitol, but not by diisopropyl fluorophosphate or benzamidine.