Fluorescence measurements of environmental relaxation at the lipid-water interface region of bilayer membranes

Abstract

Nanosecond time-resolved emission spectroscopy is used to characterize the complex fluorescence behavior of the probe 2-p-toluidinonaphthalene 6-sulfonate (2,6 p-TNS) when adsorbed to several bilayer membrane system. These include egg phosphatidylcholine vesicles with and without added cholesterol as well as erythrocyte ghost membranes. In each case a nanosecond time-dependent shift of the fluorescence emission to lower energy follows pulsed photoexcitation. The properties of the time-resolved surfaces obtained are consistent with a non-exponential decay law which describes a continuous interaction process of 2,6 p-TNS with its local environment in the membrane. This environment consists in part of polar residues (water plus polar head region) undergoing nanosecond motions. The pure phosphatidylcholine bilayer system was studied at four temperatures and electronic and spectral relaxation contributions to the total fluorescence decay were separated. Temperature coefficients for empirical rate parameters derived for the separated processes were obtained. It appears that a treatment of the fluorescence behavior of amphiphilic probes such as 2,6 p-TNS adsorbed to bilayer membranes at temperatures near ambient in which a single lifetime and radiative decay channel have been assumed is inappropriate.