Exclusion of a cholesterol analog from the cholesterol-rich phase in model membranes.
Vesicles of phosphatidylcholine/cholesterol mixtures show a wide composition range with coexistence of two fluid phases, the 'liquid disordered' (cholesterol-poor) and 'liquid ordered' (cholesterol-rich) phases. These systems have been widely used as models of membranes exhibiting lateral heterogeneity (membrane domains). The distributions of two fluorescent probes (a fluorescent cholesterol analog, NBD-cholesterol, and a lipophilic rhodamine probe, octadecylrhodamine B) in dimyristoylphosphatidylcholine/cholesterol vesicles were studied, at 30 degrees C and 40 degrees C. The steady-state fluorescence intensity of both probes decreases markedly with increasing cholesterol concentration, unlike the fluorescence lifetimes. The liquid ordered to liquid disordered phase partition coefficients K(p) were measured, and values much less than unity were obtained for both probes, pointing to preference for the cholesterol-poor phase. Globally analyzed time-resolved energy transfer results confirmed these findings. It is concluded that, in particular, NBD-cholesterol is not a suitable cholesterol analog and its distribution behavior in phosphatidylcholine/cholesterol bilayers is in fact opposite to that of cholesterol.
Loura
L M
LM
Centro de Quimica-Fisica Molecular, Instituto Superior Tecnio, P-1049-001 Lisbon, Portugal.
Fedorov
A
A
Prieto
M
M
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives / Q000737:chemistry*; D002784:Cholesterol / Q000031:analogs & derivatives / Q000737:chemistry*; D005456:Fluorescent Dyes; D008051:Lipid Bilayers / Q000737:chemistry*; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D012235:Rhodamines; D013696:Temperature
D005456:Fluorescent Dyes; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D012235:Rhodamines; C425723:octadecylrhodamine B; C035784:N(1)-cholesterylcarbamoyl-N(8)-(4-nitrobenzo-2-oxa-1,3-diazole)-3,6-dioxaoctyl-1,8-diamine; D002784:Cholesterol; D009327:4-Chloro-7-nitrobenzofurazan