Figure 3

Thin-layer chromatography (TLC) demonstrates phospholipase A2 activity of Plp. BODIPY-labeled phosphatidylcholine (BPC) was incubated with various standard enzymes or sample preparations for 1 h at 37°C. Subsequently, the lipids were extracted and separated by TLC. (A) The cleavage patterns of BPC by standard proteins PLA2, PLC, and PLD were able to distinguish the different phospholipase activities. (B) Cleavage patterns of BPC by supernatants (lanes 2 and 3) and cell lysates (lanes 4 and 5) from E. coli DH5α containing cloned plp (lanes 3 and 5) or just the cloning vector pSUP202 (lanes 2 and 4). Lane 1 contains only BPC incubated in the presence of PBS buffer. BLPC, BODIPY-labeled lysophosphatidylcholine; PA, phosphatic alcohol; PBt, phosphaticbutanol; DAG, di-acylglycerol.