Cu²⁺-Directed Liposome Membrane Fusion, Positive-Stain Electron Microscopy, and Oxidation

Natural lipid headgroups contain a few types of metal ligands, such as phosphate, amine, and serine, which interact with metal ions differently. Herein, we studied the binding between Cu²⁺ and liposomes with four types of headgroups: phosphocholine (PC), phosphoglycerol (PG), phosphoserine (PS), and cholinephosphate (CP). Using fluorescently headgroup-labeled liposomes, Cu²⁺ strongly quenched the CP and PS liposomes, whereas quenching of PC and PG was weaker. Dynamic light scattering indicated that all of the four liposomes aggregated at high Cu²⁺ concentrations, and ethylenediaminetetraacetic acid (EDTA) only restored the original size of the PC liposome, implying fusion of the other three types of liposomes. The leakage tests revealed that the integrity of PC liposomes was not affected by Cu²⁺, but the other three liposomes leaked. Under TEM, all of the liposomes show a positive-stain feature in the presence of Cu²⁺ and Cu²⁺-stained individual liposomes with a short incubation time (<1 min). The oxidative catalytic property of Cu²⁺ was also tested, and a tight binding by the PS liposome inhibited the activity of Cu²⁺. Finally, a model of interaction for each liposome was proposed, and each one has a different metal-binding and interaction mechanism.