Phospholipases A(1) and A(2) frequently coexist in biological systems. Generation of lysophosphatidylcholine (LPC) in such systems cannot be assigned to any of these types of enzymes unless the position of the fatty acid in the lysocompound can be unambiguously determined. We here present a simple method to achieve this purpose. It is based on the initial chemical acylation of the isolated LPC with a labeled fatty acid, followed by the enzymatic analysis of the resulting phosphatidylcholine (PC), using snake or bee venom phospholipase A(2). Thus, if treatment of the PC with this enzyme releases a labeled free fatty acid, it is demonstrated that the initial LPC was acylated at position sn-1, whereas if the product of hydrolysis yields labeled LPC, then the initial LPC was acylated at position sn-2. This is the first method devised to determine the source of LPC in the presence of mixtures of phospholipases A(1) and A(2) in complex biological systems.
Copyright 1999 Academic Press.