Lys-49-phospholipases A2 as active enzyme for β-arachidonoyl phospholipid bilayer membranes

Abstract

Phospholipases A2 containing Lys-49 have been reported to be extremely weak or inactive as enzyme. We have recently shown that basic proteins I and II (BP-I and BP-II), Lys- 49-PLA2s isolated from the venom of Trimeresurus flavoviridis (Habu snake), are potent to hydrolyze the arachidonate of 2-arachidonoyl-l-stearoyl-L-3-phosphatidylcholine (ASPC) in bilayer vesicles. In order to ensure such enzymatic activity of Lys-49-PLA2s, two other Lys-49- PLA2s from different snake venoms, myotoxin II (from Bothrops asper) and App-K49 (form Agkistrodon piscivorus piscivorus), were examined. Myotoxin II was found to be very active, even more potent than BP-II, liberating about 80% of arachidonic acid from liposomes. App-K49 was also active (about 50%) for ASPC liposomes. They were very weak or almost inactive for ASPC micelles and monomers. All these Lys-49-PLA2s were inactive for ASPC liposomes in the absence of Ca 2+. These results clearly demonstrated that Lys-49-PLA2s are the enzymes to hydrolyze the C2-ester bond of ASPC in bilayer membranes.