Micromolar concentrations of Zn2+ potentiates Ca2+-induced phase separation of phosphatidyl serine containing...

Fluorescence quenching of 1-acyl-2-[6[(7 nitro-2,1,3-benzoxadiazol-4yl) amino]caproyl] phosphatidyl choline in small unilamellar vesicles consisting of phosphatidyl serine has been used to monitor the lipid phase separation induced by Zn2+ and Ca2+. Phase separation of vesicle membranes was observed with Zn2+ at concentrations as low as 125 microM. Low concentrations of Zn2+ required long incubation times to reach maximal quenching (120 minutes at 375 microM). When low concentrations of Ca2+ were added to the preparation during the developing phase of Zn2+-induced quenching, an explosive increase in fluorescence quenching was instantenously observed. Phase separation induced by sub-millimolar concentrations of Ca2+ could be increased at least 4 times when vesicles were pre-incubated with 250 microM of Zn2+.