Fig. 3.

Degradation of truncated ox-PCs by LPLA2. The reaction mixture contained 49 mM sodium citrate (pH 4.5), 10 μg/ml BSA, liposomes (130 μM PL), and 40 ng/ml of recombinant mouse LPLA2 in 500 μl of total volume. The liposomes consisted of DODPC/truncated ox-PC/NAS (2.4:1:1.05, molar ratio). The reaction was initiated by the addition of recombinant LPLA2 and kept for 30 to 120 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) (upper panels) or chloroform/methanol/water (60:35:8, v/v) (lower panels) as described in the Materials and Methods. 1-O-Pal-NAS, 1-O-palmitoyl-NAS; PA, palmitic acid; LysoPC, 1-palmitoyl-sn-glycero-3-phosphocholine. The truncated ox-PC tested was POVPC, PGPC, PONPC, and PAzPC in A, B, C, and D, respectively.