Extended Data Fig. 4: PLA2G15 has high catalytic activity towards BMP under optimized enzyme reaction conditions.

a, BMP hydrolase activity of PLA2G15 has an acidic pH optimum. PLA2G15-6xHis was incubated with 3,3’ 18:1-S,S BMP under the indicated pH conditions. A representative graph is shown for an experiment repeated at least 3 times. Data are mean ± s.d. of n = 3 independent replicates. b, PLA2G15−6xHis was incubated with 1 µM of 3,3’ 18:1-S,S BMP and at different time points and enzyme concentrations. A representative graph indicates linearity in increments across multiple enzyme concentrations at a 30 s time point. c, Standard curves generated from measuring the indicated LPG and GPG standards. d, Standard curves generated from measuring the indicated BMP standards. e-h, PLA2G15 deacylates BMP to LPG with high catalytic efficiency that is comparable to that against PG phospholipid and this activity correlates with acyl chain length. Recombinant PLA2G15 was incubated with the indicated phospholipids under acidic conditions (pH = 5.0) for 30 s and reactions were stopped by heating. Phospholipids were incorporated in liposomes with non-cleavable 18:1 diether PC and indicated LPG intermediates were measured. Each experiment was repeated at least three times, and a representative graph is shown. K_cat/K_m is a measure of kinetic efficiency. Data are mean ± s.d. of n = 3 independent replicates. i-l, PLA2G15 deacylates BMP to GPG with high efficiency. Recombinant PLA2G15 was incubated with the indicated phospholipids under acidic conditions (pH = 5.0) for 30 s and reactions were stopped by heating. Phospholipids were incorporated in liposomes with non-cleavable 18:1 diether PC and GPG product was measured. Each experiment was repeated at least three times, and a representative graph is shown. “app” is defined as apparent because the catalytic efficiency is derived from a two-step reaction. Data are mean ± s.d. of n = 3 independent replicates.

Source data