Phase-dependent lateral diffusion of α-tocopherol in DPPC liposomes monitored by fluorescence quenching

The temperature-dependent fluorescence quenching of an amphiphilic palmitoyl derivative of 2,3-diazabicyclo[2.2.2]oct-2-ene (Fluorazophore-L) by α-tocopherol (α-Toc) has been determined in liposomes composed of a saturated lipid, 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). The mutual lateral diffusion coefficients (D(L)) were extracted according to a laterally diffusion-controlled dynamic quenching model. Three distinct temperature regimes were identified: one between 65 and 39 °C, where the lateral diffusion coefficients were in the range of 10(-7) cm(2) s(-1) and the lifetime of the probe was monoexponential in the absence of α-Toc, a second one between 39 and 30 °C, where the lateral diffusion coefficients were in the range of 10(-8) cm(2) s(-1) and the lifetime of the probe was biexponential in the absence of α-Toc, and a third one below 30 °C, in which no diffusion was detectable, suggesting D(L) < 10(-9) cm(2)s (-1). These temperature domains were assigned, supported by differential scanning calorimetry (DSC) measurements, to the liquid-crystalline, ripple, and solid-gel phases of DPPC liposomes in the presence of the two additives. The absolute values of the individual lateral diffusion coefficients (taken as (1)/(2) of the D(L) values) of the Fluorazophore-L/α-Toc (ca. 2.5 × 10(-7) cm(2) s(-1) at 52 °C) couple demonstrates that α-Toc does not diffuse at an unexpectedly high rate in comparison to the self-diffusion of DPPC (1.5 × 10(-7) cm(2) s(-1) at 52 °C). However, diffusion in DPPC liposomes is distinctly slower than that in POPC ones (e.g., D(L) = 4.9 × 10(-7) cm(2) s(-1) versus 6.4 × 10(-7) cm(2) s(-1) at 50 °C), with an activation energy of 49 ± 5 kJ mol(-1) (value for POPC: 47 ± 5 kJ mol(-1)), in the temperature range of the liquid-crystalline phase. Diffusion in the ripple phase, that is, below the main phase transition temperature, was found to be non-negligible, with an apparent activation energy of 175 ± 50 kJ mol(-1).