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Biophys J, January 2000, p. 267-280, Vol. 78, No. 1

Kinetics of Amphiphile Association with Two-Phase Lipid Bilayer Vesicles

Antje Pokorny,* Paulo F. F. Almeida,* Eurico C. C. Melo,dagger and Winchil L. C. Vaz*

 *Universidade de Coimbra, Departamento de Química, 3049 Coimbra Codex, and  dagger Instituto de Tecnologia Química e Biológica, 2780 Oeiras, Portugal

We examined the consequences of membrane heterogeneity for the association of a simple amphiphilic molecule with phospholipid vesicles with solid-liquid and liquid-liquid phase coexistence. To address this problem we studied the association of a single-chain, fluorescent amphiphile with dimyristoylphosphatidylcholine (DMPC) vesicles containing varying amounts of cholesterol. DMPC bilayers containing 15 mol% cholesterol show a region of solid-liquid-ordered (s-ell o) coexistence below the Tm of pure DMPC (23.9°C) and a region of liquid-disordered-liquid-ordered coexistence (ell d-ell o) above the Tm. We first examined equilibrium binding and kinetics of amphiphile insertion into single-phase vesicles (s, ell d, and ell o phase). The data obtained were then used to predict the behavior of the equivalent process in a two-phase system, taking into account the fractions of phases present. Next, the predicted kinetics were compared to experimental kinetics obtained from a two-phase system. We found that association of the amphiphile with lipid vesicles is not influenced by the existence of ell d-ell o phase boundaries but occurs much more slowly in the s-ell o phase coexistence region than expected on the basis of phase composition.

Biophys J, January 2000, p. 267-280, Vol. 78, No. 1
© 2000 by the Biophysical Society   0006-3495/00/01/267/14  $2.00


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