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Departamento de Quimica, Universidade de Coimbra, Coimbra, Portugal
Correspondence: Address reprint requests to Prof. Winchil L. C. Vaz, Departamento de Quimica, Universidade de Coimbra, 3004-535 Coimbra, Portugal. Tel.: 351-23-982-4861; Fax: 351-23-982-7703; E-mail: wvaz{at}ci.uc.pt.
We have measured the rates of insertion into, desorption from, and spontaneous interlayer translocation (flip-flop) in liquid-disordered and liquid-ordered phase lipid bilayer membranes, of the fluorescent phospholipid derivative NBD-dimyristoylphosphatidyl ethanolamine. This study made use of a recently described method that exploits a detailed knowledge of the binding kinetics of an amphiphile to bovine serum albumin, to recover the insertion and desorption rate constants when the albumin-bound amphiphile is transferred through the aqueous phase to the membrane and vice versa. The lipid bilayers, studied as large unilamellar vesicles, were prepared from pure 1-palmitoyl-2-oleoylphosphatidylcholine in the liquid-disordered phase; and from two cholesterol-containing binary lipid mixtures, 1-palmitoyl-2-oleoylphosphatidylcholine and cholesterol (molar ratio of 1:1), and egg sphingomyelin and cholesterol (molar ratio of 6:4), both in the liquid-ordered phase. Insertion, desorption, and translocation rate constants and equilibrium constants for association of the amphiphile monomer with the lipid bilayers were directly measured between 15° and 35°C, and the standard free energies, enthalpies, and entropies, as well as the activation energies for these processes, were derived from this data. The equilibrium partition coefficients for partitioning of the amphiphile between the aqueous phase and the different membrane phases were also derived, and permitted the estimation of hypothetical partition coefficients and the respective energetic parameters for partitioning between the different lipid phases if these were to coexist in the same membrane.
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M. J. Moreno, L. M. B. B. Estronca, and W. L. C. Vaz Translocation of Phospholipids and Dithionite Permeability in Liquid-Ordered and Liquid-Disordered Membranes Biophys. J., August 1, 2006; 91(3): 873 - 881. [Abstract] [Full Text] [PDF]
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J. L. Sampaio, M. J. Moreno, and W. L. C. Vaz Kinetics and Thermodynamics of Association of a Fluorescent Lysophospholipid Derivative with Lipid Bilayers in Liquid-Ordered and Liquid-Disordered Phases Biophys. J., June 1, 2005; 88(6): 4064 - 4071. [Abstract] [Full Text] [PDF]
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L. M. B. B. Estronca, M. J. Moreno, J. A. N. Laranjinha, L. M. Almeida, and W. L. C. Vaz Kinetics and Thermodynamics of Lipid Amphiphile Exchange between Lipoproteins and Albumin in Serum Biophys. J., January 1, 2005; 88(1): 557 - 565. [Abstract] [Full Text] [PDF]
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