Miscibility and phase behavior of N-acylethanolamine/ diacylphosphatidylethanolamine binary mixtures of matched acyl chainlengths (n = 14, 16)
Ravi Kanth Kamlekar 1, S. Satyanarayana 1, Derek Marsh 2 and Musti J. Swamy 1*
1 University of Hyderabad
2 Max-Planck Instituet fuer biophys. Chemie
* To whom correspondence should be addressed. E-mail: mjssc{at}uohyd.ernet.in.
Submitted on September 2, 2006
Revised on October 9, 2006
Accepted on 24 January 2007
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Abstract |
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The miscibility and phase behavior of hydrated binary mixtures of two N-acylethanolamines (NAEs), N-myristoylethanolamine (NMEA) and N-palmitoylethanolamine (NPEA), with the corresponding diacyl phosphatidylethanolamines (PEs), dimyristoylphosphatidylethanolamine (DMPE) and dipalmitoylphosphatidylethanolamine (DPPE), respectively, have been investigated by differential scanning calorimetry (DSC), spin-label ESR and 31P-NMR spectroscopy. Temperature-composition phase diagrams for both NMEA/DMPE and NPEA/DPPE binary systems were established from high sensitivity DSC. The structures of the phases involved were determined by 31P-NMR spectroscopy. For both systems, complete miscibility in the fluid and gel phases is indicated by DSC and ESR, up to 35 mol% of NMEA in DMPE and 40 mol% of NPEA in DPPE. At higher contents of the NAEs, extensive solid-fluid phase separation and solid-solid immiscibility occur depending on the temperature. Characterization of the structures of the mixtures formed with 31P-NMR spectroscopy shows that up to 75 mol% of NAE, both DMPE and DPPE form lamellar structures in the gel phase as well as up to at least 65° in the fluid phase. ESR spectra of phosphatidylcholine spin-labeled at C-5 position in the sn-2 acyl chain present at a probe concentration of 1 mol% exhibit strong spin-spin broadening in the low-temperature region for both systems, suggesting that the acyl chains pack very tightly and exclude the spin label. However, spectra recorded in the fluid phase do not exhibit any spin-spin broadening, and indicate complete miscibility of the two components. The miscibility of NAE and diacyl PE of matched chainlengths is significantly less than that found earlier for NPEA and dipalmitoylphosphatidylcholine, an observation that is consistent with the notion that the NAEs are most likely stored as their precursor lipids (N-acyl PEs) and are generated only when the system is subjected to membrane stress.
Key Words:
31P-NMR spectroscopy, differential scanning calorimetry, lipid membrane, phase diagram, phase transition, spin-label ESR spectroscopy
