Fluorescence spectroscopy of an oriented model membrane

Abstract

We have devised a simple method that makes it feasible to apply fluorescence techniques to lipid bilayer membranes to elucidate aspects of their structure and dynamics. Fluorescence excitation, emission, and polarization spectra were obtained from a single spherical bilayer membrane consisting of oxidized cholesterol and fluorescent probe. The emission transition moments of N,N'-di(octadecyl)oxacarbocyanine and 12-(9-anthroyl)-stearic acid were found to be aligned parallel to the plane of the bilayer, whereas that of p-bis-[2-(4-methyl-5-phenyloxazolyl)]-benzene was aligned in a perpendicular direction. All three probes exhibited appreciable rotational mobility, parallel to the plane of the bilayer, in durations of nanoseconds. An attractive feature of this model membrane is that fluorescence measurements can be made at the same time as electrical measurements and perturbations. Also, it may be possible to incorporate functional protein assemblies into this model and to use fluorescence spectroscopy to delineate some aspects of their assembly and function.