Dimyristoylphosphatidylcholine/C16 : 0-ceramide binary liposomes studied by differential scanning calorimetry and wide- and small-angle X-ray scattering

J. M. Holopainen, Jesper Lemmich, F. Richter, Ole G. Mouritsen, G. Rapp, P. K. J. Kinnunen

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Abstract

Ceramide has recently been established as a central messenger in the signaling cascades controlling cell behavior. Physicochemical studies have revealed a strong tendency of this lipid toward phase separation in mixtures with phosphatidylcholines. The thermal phase behavior and structure of fully hydrated binary membranes composed of dimyristoylphosphatidylcholine (DMPC) and N-palmitoyl-ceramide (C16:0-ceramide, up to a mole fraction X-cer = 0.35) were resolved in further detail by high-sensitivity differential scanning calorimetry (DSC) and x-ray diffraction. Both methods reveal very strong hysteresis in the thermal phase behavior of ceramide-containing membranes. A partial phase diagram was constructed based on results from a combination of these two methods. DSC heating scans show that with increased X-cer the pretransition temperature T-P first increases, whereafter at X-cer > 0.06 it can no longer be resolved. The main transition enthalpy Delta H remains practically unaltered while its width increases significantly, and the upper phase boundary temperature of the mixture shifts to similar to 63 degrees C at X-cer = 0.30. Upon cooling, profound phase separation is evident, and for all of the studied compositions there is an endotherm in the region close to the T-m for DMPC. At X-cer greater than or equal to 0.03 a second endotherm is evident at higher temperatures, starting at 32.1 degrees C and reaching 54.6 degrees C at X-cer = 0.30. X-ray small-angle reflection heating scans reveal a lamellar phase within the temperature range of 15-60 degrees C, regardless of composition. The pretransition is observed up to X-cer < 0.18, together with an increase in T-p. In the gel phase the lamellar repeat distance d increases from similar to 61 Angstrom at X-cer = 0.03, to 67 Angstrom at X-cer = 0.35. In the fluid phase increasing X-cer from 0.06 to 0.35 augments d from 61 Angstrom to 64 Angstrom. An L-beta/L-alpha (ripple/fluid) phase coexistence region is observed at high temperatures (from 31 to 56.5 degrees C) when X-cer > 0.03. With cooling from temperatures above 50 degrees C we observe a slow increase in d as the coexistence region is entered. A sudden solidification into a metastable, modulated gel phase with high d values is observed for all compositions at similar to 24 degrees C. The anomalous swelling for up to X-cer = 0.30 in the transition region is interpreted as an indication of bilayer softening and thermally reduced bending rigidity.
Original languageEnglish
JournalBiophys J.
Volume78
Issue number5
Pages (from-to)2459-2469
Publication statusPublished - 2000

Cite this

Holopainen, J. M., Lemmich, J., Richter, F., Mouritsen, O. G., Rapp, G., & Kinnunen, P. K. J. (2000). Dimyristoylphosphatidylcholine/C16 : 0-ceramide binary liposomes studied by differential scanning calorimetry and wide- and small-angle X-ray scattering. Biophys J., 78(5), 2459-2469.