Fig. 2.

Phospholipase C activity of PLC-γ1 (D1165H) in lipid vesicles. (a) XY-69 (0.5 μM) was embedded into lipid vesicles containing PE (200 μM) and PIP2 (20 μM) and added to PLC-γ1 (D1165H) with indicated concentration to initiate the hydrolysis. The reaction progression at 20 °C was monitored continuously for 60 min by fluorescence (λex/λem = 485/520 nm). The final assay buffer has a pH of 7.4 and free Ca2+ concentration at 390 nM. (b) The concentration of PE and PIP2 in lipid vesicles was 192 and 48 μM, respectively. The experiment was run similarly as described in (a)