Vesicles of phosphatidylcholine/cholesterol mixtures show a wide compositio
n range with coexistence of two fluid phases, the 'liquid disordered' (chol
esterol-poor) and 'liquid ordered' (cholesterol-rich) phases. These systems
have been widely used as models of membranes exhibiting lateral heterogene
ity (membrane domains). The distributions of two fluorescent probes (a fluo
rescent cholesterol analog. NBD-cholesterol, and a lipophilic rhodamine pro
be, octadecylrhodamine B) in dimyristoylphosphatidylcholine/cholesterol ves
icles were studied, at 30 degreesC and 40 degreesC. The steady-state fluore
scence intensity of both probes decreases markedly with increasing choleste
rol concentration. unlike the fluorescence lifetimes. The liquid ordered to
liquid disordered phase partition coefficients K-p were measured, and valu
es much less than unity were obtained for both probes, pointing to preferen
ce for the cholesterol-poor phase. Globally analyzed time-resolved energy t
ransfer results confirmed these findings. It is concluded that, in particul
ar. NBD-cholesterol is not a suitable cholesterol analog and its distributi
on behavior in phosphatidylcholine/cholesterol bilayers is in fact opposite
to that of cholesterol. (C) 2001 Elsevier Science B.V. All rights reserved
.