Fluorescence lifetime distributions of 1,6-diphenyl-1,3,5-hexatriene reveal the effect of cholesterol on the microheterogeneity of erythrocyte membrane

Abstract

The fluorescence decay of 1,6 diphenyl-1,3,5-hexatriene (DPH) has been used to characterize aspects of the erythrocyte membrane structure related to the microheterogeneity of the lipid bilayer. The DPH decay has been studied using frequency domain fluorometry and the data analyzed either by a model of discrete exponential components or a model that assumes a continuous distribution of lifetime values. The main intensity fraction was associated with a lifetime value centered at about 11 ns in the erythrocyte membrane, but a short component of very low fractional intensity had to be considered to obtain a good fit to the data. The lifetime value of the long component was insensitive to temperature, while the width of the distribution decreased with increasing temperature. In multilamellar liposomes prepared from phospholipids extracted from the erythrocytes, the long lifetime component showed a temperature dependence. The depletion of 27% of the cholesterol in the erythrocyte membrane induced a broadening of the distribution, suggesting a homogenizing effect of cholesterol. This effect has also been detected in egg phosphatidylcholine at a very low cholesterol/phospholipid molar ratio. The role of cholesterol on membrane heterogeneity is discussed in relation to the effect of cholesterol on water penetration.