Skip to main content
Biophysical Journal logoLink to Biophysical Journal
. 1995 Mar;68(3):978–987. doi: 10.1016/S0006-3495(95)80273-0

Liquid-crystalline phases of cholesterol/lipid bilayers as revealed by the fluorescence of trans-parinaric acid.

C Reyes Mateo 1, A Ulises Acuña 1, J C Brochon 1
PMCID: PMC1281821  PMID: 7756560

Abstract

The presence of two liquid-crystalline phases, alpha and beta, in mixed bilayers of dimyristoylphosphatidylcholine/cholesterol was detected by the changes in the distribution of the fluorescence lifetimes of t-PnA, as analyzed by the Maximum Entropy Method. The formation of the liquid-ordered beta-phase, in the 30-40 degrees C temperature range as a function of cholesterol concentration (0-40 mol%), could be related quantitatively to the relative amplitude of a long lifetime component of the probe (10-14 ns). Based on this evidence, the phase behavior of mixtures of the unsaturated lipid palmitoyloleoylphosphatidylcholine and cholesterol was determined using the same technique, for cholesterol concentrations in the 0-50 mol% range, between 10 and 40 degrees C. It was found that two liquid-crystalline phases are also formed in this system, with physical properties reminiscent of the alpha- and beta-phases formed with saturated lipids. However, in this case it was determined that, for temperatures in the physiological range, the alpha- and beta-phases coexist up to 40 mol% cholesterol. This finding may be of significant biological relevance, because it supports the long held notion that cholesterol is responsible for the lipid packing heterogeneity of several natural membranes rich in unsaturated lipid components.

Full text

PDF
978

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Almeida P. F., Vaz W. L., Thompson T. E. Lateral diffusion in the liquid phases of dimyristoylphosphatidylcholine/cholesterol lipid bilayers: a free volume analysis. Biochemistry. 1992 Jul 28;31(29):6739–6747. doi: 10.1021/bi00144a013. [DOI] [PubMed] [Google Scholar]
  2. Bloom M., Evans E., Mouritsen O. G. Physical properties of the fluid lipid-bilayer component of cell membranes: a perspective. Q Rev Biophys. 1991 Aug;24(3):293–397. doi: 10.1017/s0033583500003735. [DOI] [PubMed] [Google Scholar]
  3. Brochon J. C. Maximum entropy method of data analysis in time-resolved spectroscopy. Methods Enzymol. 1994;240:262–311. doi: 10.1016/s0076-6879(94)40052-0. [DOI] [PubMed] [Google Scholar]
  4. Cross A. J., Fleming G. R. Analysis of time-resolved fluorescence anisotropy decays. Biophys J. 1984 Jul;46(1):45–56. doi: 10.1016/S0006-3495(84)83997-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gallay J., Vincent M. Cardiolipin-cholesterol interactions in the liquid-crystalline phase: a steady-state and time-resolved fluorescence anisotropy study with cis- and trans-parinaric acids as probes. Biochemistry. 1986 May 6;25(9):2650–2656. doi: 10.1021/bi00357a054. [DOI] [PubMed] [Google Scholar]
  6. Gordon L. M., Mobley P. W., Esgate J. A., Hofmann G., Whetton A. D., Houslay M. D. Thermotropic lipid phase separations in human platelet and rat liver plasma membranes. J Membr Biol. 1983;76(2):139–149. doi: 10.1007/BF02000614. [DOI] [PubMed] [Google Scholar]
  7. Ipsen J. H., Karlström G., Mouritsen O. G., Wennerström H., Zuckermann M. J. Phase equilibria in the phosphatidylcholine-cholesterol system. Biochim Biophys Acta. 1987 Nov 27;905(1):162–172. doi: 10.1016/0005-2736(87)90020-4. [DOI] [PubMed] [Google Scholar]
  8. Ipsen J. H., Mouritsen O. G., Bloom M. Relationships between lipid membrane area, hydrophobic thickness, and acyl-chain orientational order. The effects of cholesterol. Biophys J. 1990 Mar;57(3):405–412. doi: 10.1016/S0006-3495(90)82557-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Lentz B. R., Barrow D. A., Hoechli M. Cholesterol-phosphatidylcholine interactions in multilamellar vesicles. Biochemistry. 1980 Apr 29;19(9):1943–1954. doi: 10.1021/bi00550a034. [DOI] [PubMed] [Google Scholar]
  10. Livesey A. K., Brochon J. C. Analyzing the distribution of decay constants in pulse-fluorimetry using the maximum entropy method. Biophys J. 1987 Nov;52(5):693–706. doi: 10.1016/S0006-3495(87)83264-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ludescher R. D., Peting L., Hudson S., Hudson B. Time-resolved fluorescence anisotropy for systems with lifetime and dynamic heterogeneity. Biophys Chem. 1987 Oct;28(1):59–75. doi: 10.1016/0301-4622(87)80075-3. [DOI] [PubMed] [Google Scholar]
  12. Mabrey S., Mateo P. L., Sturtevant J. M. High-sensitivity scanning calorimetric study of mixtures of cholesterol with dimyristoyl- and dipalmitoylphosphatidylcholines. Biochemistry. 1978 Jun 13;17(12):2464–2468. doi: 10.1021/bi00605a034. [DOI] [PubMed] [Google Scholar]
  13. Mabrey S., Sturtevant J. M. Investigation of phase transitions of lipids and lipid mixtures by sensitivity differential scanning calorimetry. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3862–3866. doi: 10.1073/pnas.73.11.3862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mateo C. R., Lillo M. P., González-Rodríguez J., Acuña A. U. Lateral heterogeneity in human platelet plasma membrane and lipids from the time-resolved fluorescence of trans-parinaric acid. Eur Biophys J. 1991;20(1):53–59. doi: 10.1007/BF00183279. [DOI] [PubMed] [Google Scholar]
  15. Mortensen K., Pfeiffer W., Sackmann E., Knoll W. Structural properties of a phosphatidylcholine-cholesterol system as studied by small-angle neutron scattering: ripple structure and phase diagram. Biochim Biophys Acta. 1988 Nov 22;945(2):221–245. doi: 10.1016/0005-2736(88)90485-3. [DOI] [PubMed] [Google Scholar]
  16. Parasassi T., Di Stefano M., Loiero M., Ravagnan G., Gratton E. Influence of cholesterol on phospholipid bilayers phase domains as detected by Laurdan fluorescence. Biophys J. 1994 Jan;66(1):120–132. doi: 10.1016/S0006-3495(94)80763-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pasenkiewicz-Gierula M., Subczynski W. K., Kusumi A. Rotational diffusion of a steroid molecule in phosphatidylcholine-cholesterol membranes: fluid-phase microimmiscibility in unsaturated phosphatidylcholine-cholesterol membranes. Biochemistry. 1990 May 1;29(17):4059–4069. doi: 10.1021/bi00469a006. [DOI] [PubMed] [Google Scholar]
  18. Recktenwald D. J., McConnell H. M. Phase equilibria in binary mixtures of phosphatidylcholine and cholesterol. Biochemistry. 1981 Jul 21;20(15):4505–4510. doi: 10.1021/bi00518a042. [DOI] [PubMed] [Google Scholar]
  19. Reinl H., Brumm T., Bayerl T. M. Changes of the physical properties of the liquid-ordered phase with temperature in binary mixtures of DPPC with cholesterol: A H-NMR, FT-IR, DSC, and neutron scattering study. Biophys J. 1992 Apr;61(4):1025–1035. doi: 10.1016/S0006-3495(92)81910-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Reyes Mateo C., Brochon J. C., Pilar Lillo M., Ulises Acuña A. Lipid clustering in bilayers detected by the fluorescence kinetics and anisotropy of trans-parinaric acid. Biophys J. 1993 Nov;65(5):2237–2247. doi: 10.1016/S0006-3495(93)81257-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Reyes Mateo C., Tauc P., Brochon J. C. Pressure effects on the physical properties of lipid bilayers detected by trans-parinaric acid fluorescence decay. Biophys J. 1993 Nov;65(5):2248–2260. doi: 10.1016/S0006-3495(93)81258-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ruggiero A., Hudson B. Analysis of the anisotropy decay of trans-parinaric acid in lipid bilayers. Biophys J. 1989 Jun;55(6):1125–1135. doi: 10.1016/S0006-3495(89)82909-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ruggiero A., Hudson B. Critical density fluctuations in lipid bilayers detected by fluorescence lifetime heterogeneity. Biophys J. 1989 Jun;55(6):1111–1124. doi: 10.1016/S0006-3495(89)82908-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sankaram M. B., Thompson T. E. Cholesterol-induced fluid-phase immiscibility in membranes. Proc Natl Acad Sci U S A. 1991 Oct 1;88(19):8686–8690. doi: 10.1073/pnas.88.19.8686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sankaram M. B., Thompson T. E. Interaction of cholesterol with various glycerophospholipids and sphingomyelin. Biochemistry. 1990 Nov 27;29(47):10670–10675. doi: 10.1021/bi00499a014. [DOI] [PubMed] [Google Scholar]
  26. Schroeder F., Jefferson J. R., Kier A. B., Knittel J., Scallen T. J., Wood W. G., Hapala I. Membrane cholesterol dynamics: cholesterol domains and kinetic pools. Proc Soc Exp Biol Med. 1991 Mar;196(3):235–252. doi: 10.3181/00379727-196-43185. [DOI] [PubMed] [Google Scholar]
  27. Scott H. L. Lipid-cholesterol interactions. Monte Carlo simulations and theory. Biophys J. 1991 Feb;59(2):445–455. doi: 10.1016/S0006-3495(91)82238-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sklar L. A., Hudson B. S., Simoni R. D. Conjugated polyene fatty acids as fluorescent probes: synthetic phospholipid membrane studies. Biochemistry. 1977 Mar 8;16(5):819–828. doi: 10.1021/bi00624a002. [DOI] [PubMed] [Google Scholar]
  29. Sklar L. A., Miljanich G. P., Dratz E. A. Phospholipid lateral phase separation and the partition of cis-parinaric acid and trans-parinaric acid among aqueous, solid lipid, and fluid lipid phases. Biochemistry. 1979 May 1;18(9):1707–1716. doi: 10.1021/bi00576a012. [DOI] [PubMed] [Google Scholar]
  30. Sklar L. A. The partition of cis-parinaric acid and trans-parinaric acid among aqueous, fluid lipid, and solid lipid phases. Mol Cell Biochem. 1980 Nov 20;32(3):169–177. doi: 10.1007/BF00227444. [DOI] [PubMed] [Google Scholar]
  31. Thewalt J. L., Bloom M. Phosphatidylcholine: cholesterol phase diagrams. Biophys J. 1992 Oct;63(4):1176–1181. doi: 10.1016/S0006-3495(92)81681-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vist M. R., Davis J. H. Phase equilibria of cholesterol/dipalmitoylphosphatidylcholine mixtures: 2H nuclear magnetic resonance and differential scanning calorimetry. Biochemistry. 1990 Jan 16;29(2):451–464. doi: 10.1021/bi00454a021. [DOI] [PubMed] [Google Scholar]

Articles from Biophysical Journal are provided here courtesy of The Biophysical Society

RESOURCES