Transbilayer diffusion of divalent cations into liposomes mediated by lipidic particles of phosphatidate

Liposomes formed from egg-yolk phosphatidylcholine:egg-yolk phosphatidate (molar ratio 2:1) containing pBR322 DNA and DNase I were induced to form, with divalent cations, bilayer/nonbilayer phase transitions of phosphatidate which allowed cation diffusion into liposomes; then cation diffusion was measured by the activation of the hydrolysis of DNase I on DNA. The formation of phosphatidate transitions on liposomes was demonstrated by freeze-fracture and ³¹P NMR, and a direct correlation between the formation of phosphatidate transitions and the transbilayer diffusion of cations was found: only Ca²⁺ and Mn²⁺, which induce phase transitions, were able to penetrate liposomes and triggered the DNase I activity; in addition, Ca²⁺ at higher concentrations (10 mM) caused fusion of liposomes, whereas Mn²⁺ did not, suggesting that transitions induced by Mn²⁺ participated only in the diffusion of this ion; furthermore, Mg²⁺ neither formed phase transitions nor triggered the enzymatic activity. The liposomes studied represent more dynamic structures that can form phosphatidate structures involved in both (1) the interchange of divalent cations with the surroundings, thereby modulating encapsulated enzymes, and (2) the fusion of lipid vesicles probably implicated in the enrichment of liposomal content in the early Precambian Earth.