A long-lifetime Ru(II) metal-ligand complex as a membrane probe

A luminescent metal-ligand complex, [Ru(bpy)2(dppz)]2+, (where dppz is dipyrido[3,2-a:2',3'-c] phenazine), was used as a photoluminescence probe for investigating submicrosecond lipid dynamics in a dipalmitoyl-L-alpha-phosphotidylglycerol (DPPG) model bilayer system. The luminescence of [Ru(bpy)2(dppz)]2+ in buffer is completely quenched but becomes luminescent when intercalated into DPPG vesicles. The experimental results show that the emission intensity of [Ru(bpy)2(dppz)]2+ intercalated into DPPG vesicles increases dramatically as temperature is increased towards the lipid phase transition temperature. This effect is abolished in bilayers containing a high concentration (> 30 mol%) of cholesterol, suggesting this probe is sensitive to the membrane composition. Frequency-domain emission intensity decays, measured as a function of increasing temperature towards the lipid phase transition temperature (2 to 57 degrees C), display two major lifetime components. The short lifetime disappears at temperatures well above the phase transition temperature. A comparison of oxygen quenching with iodide quenching suggests the heterogeneity of probe location at temperatures well below the lipid phase transition temperature and the homogeneity of probe location at temperature well above the lipid phase transition temperature. [Ru(bpy)2(dppz)]2+ displays polarized emission, enabling the study of membrane dynamics. The long decay time displayed by this probe allows measurement of the overall rotational correlation time of lipid vesicles on the microsecond time-scale. Because of the long lifetime, polarized emission, and background free nature of the photoluminescence measurements, [Ru(bpy)2(dppz)]2+ has numerous applications in the biophysical studies of membranes.