Fig. 3.

pH dependence of transacylase activity. A: WT LPLA₂ reaction products separated by TLC, including free fatty acid (FA) and 1-O-acyl-NAS as a function of pH. B: D13F reaction products separated by TLC of as a function of pH. C: pH dependent ceramide transacylase activities of individual LPLA₂ variants. The data represent the mean ± SD (n = 3) per time point, *P < 0.05, ***P < 0.001 using a t-test. In all panels, the transacylase activity was determined by incubating DOPC/sulfatide/NAS liposomes (10/1/3 molar ratio) in 48 mM Na citrate pH 4.5 or 50 mM HEPES pH 7.4, with 30 ng of each variant for 30 min at 37°C.