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Biophys J, February 2001, p. 765-775, Vol. 80, No. 2

Interfacial Interactions of Ceramide with Dimyristoylphosphatidylcholine: Impact of the N-Acyl Chain

Juha M. Holopainen,* Howard L. Brockman,dagger Rhoderick E. Brown,dagger and Paavo K. J. Kinnunen*

 *Helsinki Biophysics and Biomembrane Group, Department of Medical Chemistry, Institute of Biomedicine, University of Helsinki, FIN-00014 Helsinki, Finland; and  dagger The Hormel Institute, University of Minnesota, Austin, Minnesota 55912 USA

The mixing behavior of dimyristoylphosphatidylcholine (DMPC) with either N-palmitoyl-sphingosine (C16:0-ceramide) or N-nervonoyl-sphingosine (C24:1-ceramide) was examined using monomolecular films. While DMPC forms highly elastic liquid-expanded monolayers, both neat C16:0-ceramide and C24:1-ceramide yield stable solid condensed monomolecular films with small areas and low interfacial elasticity. Compression isotherms of mixed C16:0-ceramide/DMPC films exhibit an apparent condensation upon increasing Xcer16:0 at all surface pressures. The average area isobars, coupled with the lack of a liquid-expanded to condensed phase transition as Xcer16:0 is increased, are indicative of immiscibility of the lipids at all surface pressures. In contrast, isobars for C24:1-ceramide/DMPC mixtures show surface pressure-dependent apparent condensation or expansion and surface pressure-area isotherms show a composition and surface pressure-dependent phase transition. This suggests miscibility, albeit non-ideal, of C24:1-ceramide and DMPC in both liquid and condensed surface phases. The above could be verified by fluorescence microscopy of the monolayers and measurements of surface potential, which revealed distinctly different domain morphologies and surface potential values for the DMPC/C16:0- and DMPC/C24:1-ceramide monolayers. Taken together, whereas C16:0-ceramide and DMPC form immiscible pseudo-compounds, C24:1-ceramide and DMPC are partially miscible in both the liquid-expanded and condensed phases, and a composition and lateral pressure-dependent two-phase region is evident between the liquid-expanded and condensed regimes. Our results provide novel understanding of the regulation of membrane properties by ceramides and raise the possibility that ceramides with different acyl groups could serve very different functions in cells, relating to their different physicochemical properties.

Biophys J, February 2001, p. 765-775, Vol. 80, No. 2
© 2001 by the Biophysical Society   0006-3495/01/02/765/11  $2.00



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