Fig. 6.

Effect of AMD on degradation of truncated ox-PC by LPLA2. The reaction mixture consisting of 50 mM sodium citrate (pH 4.5), 10 μg/ml BSA, and liposomes (131 μM PL) was preincubated with or without 9.7 μM AMD in 490 μl of total volume for 5 min at 37°C. The liposomes consisted of DODPC/DOPC or truncated ox-PC/NAS (2.4:1:1.05, molar ratio). The reaction was initiated by the addition of 10 μl of 2 μg/ml recombinant LPLA2 and kept for 60 min at 37°C. The reaction products were extracted and separated with an HPTLC plate using a solvent system consisting of chloroform/acetic acid (9:1, v/v), as described in the Materials and Methods. Formed 1-O-acyl-NAS and fatty acid were quantified as described in the Materials and Methods. Error bars indicate standard deviation (n = 3). DOPC (A), POVPC (B), PGPC (C), PONPC (D), or PAzPC (E) was loaded into DODPC-based liposomes. Cont. denotes control (without AMD).