Influence of cholesterol on equilibrium and dynamic bilayer structure of unsaturated acyl chain phosphatidylcholine vesicles as determined from higher order analysis of fluorescence anisotropy decay.
The influence of cholesterol on equilibrium and dynamic bilayer structure in minimally to highly unsaturated phosphatidylcholine (PC) vesicles has been examined by characterization of the dynamic fluorescence properties of 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1-[4-(trimethylammonio)phenyl]-6-phenyl-1,3,5-hexatriene (TMA-DPH). Large, unilamellar egg PC, palmitoyloleoyl-PC (POPC), dioleoyl-PC (DOPC), palmitoylarachidonoyl-PC (PAPC), and palmitoyldocosahexaenoyl-PC (P-22:6-PC) vesicles containing no cholesterol or approximately 15 or 30 mol % cholesterol have been examined. Equilibrium and dynamic DPH orientational properties were analyzed according to an orthogonal, bimodal orientational distribution function [Straume, M., & Litman, B.J. (1987) Biochemistry (preceding paper in this issue)]. The same mathematical formalism was applied to TMA-DPH except that probe orientational probability was permitted only in the distribution peak aligned parallel to the bilayer normal. TMA-DPH fluorescence lifetimes were consistently increased by incorporation of cholesterol into these vesicles. Greater acyl chain unsaturation and increasing temperature each promoted reduction of lifetimes in the presence or absence of cholesterol. DPH lifetimes were much less sensitive than those of TMA-DPH to changes in composition or temperature. This behavior is consistent with reduced water penetrability into liquid-crystalline bilayers as cholesterol content is increased and as acyl chain unsaturation and temperature are reduced. Cholesterol also induces substantial equilibrium ordering of the bilayer both at the hydrophobic core and at the bilayer-water interface. DPH orientational distributions were shifted in favor of alignment parallel to the acyl side chains. The distributions of both probes were narrowed in response to incorporation of cholesterol.(ABSTRACT TRUNCATED AT 250 WORDS)
Straume
M
M
Department of Biochemistry, University of Virginia School of Medicine, Charlottesville 22908.
Litman
B J
BJ
D002784:Cholesterol; D004161:Diphenylhexatriene / Q000031:analogs & derivatives; D005454:Fluorescence Polarization / Q000379:methods; D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D013329:Structure-Activity Relationship
D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D004161:Diphenylhexatriene; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene; D002784:Cholesterol