Membrane fusion mediated by phospholipase C under endosomal pH conditions

Phospholipase C (PLC) is considered to be one of key enzymes for the design of drug delivery system using the endocytosis route, because PLC can catalyze the membrane fusion between cell membranes and phospholipid vehicles (liposomes). Membrane fusion by PLC was then studied under various pHs to model the endosomal environment. The used liposomes were composed of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), and cholesterol (Ch). The membrane fusion was dominated by the enzymatic reaction at pH 6–7.5. In contrast, the membrane perturbation effect due to the conformational change of PLC could induce the membrane fusion at around pH 4. The maximal value of membrane fusion was observed at around pH 5 for three liposomes in the order of DOPC < DOPC/DPPC (1:1) < DOPC/DPPC/Ch (1:1:1). From the experiments with the hydrophobic fluorescence probe and the circular dichroic method, it was revealed that PLC took a molten-globule state, with a large fluctuation and an enzymatic activity, at around pH 4–5. The DAG-rich domain enzymatically produced by PLC played role for the field for the membrane perturbation enough to lead to the membrane fusion. The present finding would be helpful to understand the behavior of membrane fusion under the late endosomal pH condition in cell system.

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► Phospholipase C could induce the remarkable membrane fusion at endosomal pH condition.
► The above fusion resulted from the response of phospholipase C against pH-shift.
► Enzymatic production of diacylglycerol was advantageous for the interaction between liposomes.
► The membrane perturbation of liposome by phospholipase C induced membrane fusion.
► Endosomal pH gave both the enzymatic activity and the flexible structure for membrane perturbation.