Skip to main content
Biophysical Journal logoLink to Biophysical Journal
. 1997 Oct;73(4):1940–1953. doi: 10.1016/S0006-3495(97)78225-0

Molecular sorting of lipids by bacteriorhodopsin in dilauroylphosphatidylcholine/distearoylphosphatidylcholine lipid bilayers.

F Dumas 1, M M Sperotto 1, M C Lebrun 1, J F Tocanne 1, O G Mouritsen 1
PMCID: PMC1181095  PMID: 9336190

Abstract

A combined experimental and theoretical study is performed on binary dilauroylphosphatidylcholine/distearoylphosphatidylcholine (DLPC/DSPC) lipid bilayer membranes incorporating bacteriorhodopsin (BR). The system is designed to investigate the possibility that BR, via a hydrophobic matching principle related to the difference in lipid bilayer hydrophobic thickness and protein hydrophobic length, can perform molecular sorting of the lipids at the lipid-protein interface, leading to lipid specificity/selectivity that is controlled solely by physical factors. The study takes advantage of the strongly nonideal mixing behavior of the DLPC/DSPC mixture and the fact that the average lipid acyl-chain length is strongly dependent on temperature, particularly in the main phase transition region. The experiments are based on fluorescence energy transfer techniques using specifically designed lipid analogs that can probe the lipid-protein interface. The theoretical calculations exploit a microscopic molecular interaction model that embodies the hydrophobic matching as a key parameter. At low temperatures, in the gel-gel coexistence region, experimental and theoretical data consistently indicate that BR is associated with the short-chain lipid DLPC. At moderate temperatures, in the fluid-gel coexistence region, BR remains in the fluid phase, which is mainly composed of short-chain lipid DLPC, but is enriched at the interface between the fluid and gel domains. At high temperatures, in the fluid phase, BR stays in the mixed lipid phase, and the theoretical data suggest a preference of the protein for the long-chain DSPC molecules at the expense of the short-chain DLPC molecules. The combined results of the experiments and the calculations provide evidence that a molecular sorting principle is active because of hydrophobic matching and that BR exhibits physical lipid selectivity. The results are discussed in the general context of membrane organization and compartmentalization and in terms of nanometer-scale lipid-domain formation.

Full text

PDF
1940

Images in this article

Selected References

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

  1. Andersen A. S. Cell biology. Reception and transmission. Nature. 1989 Jan 5;337(6202):12–12. doi: 10.1038/337012a0. [DOI] [PubMed] [Google Scholar]
  2. Arias H. R., Sankaram M. B., Marsh D., Barrantes F. J. Effect of local anaesthetics on steroid-nicotinic acetylcholine receptor interactions in native membranes of Torpedo marmorata electric organ. Biochim Biophys Acta. 1990 Sep 7;1027(3):287–294. doi: 10.1016/0005-2736(90)90320-n. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Brown M. F. Modulation of rhodopsin function by properties of the membrane bilayer. Chem Phys Lipids. 1994 Sep 6;73(1-2):159–180. doi: 10.1016/0009-3084(94)90180-5. [DOI] [PubMed] [Google Scholar]
  5. Brumm T., Jørgensen K., Mouritsen O. G., Bayerl T. M. The effect of increasing membrane curvature on the phase transition and mixing behavior of a dimyristoyl-sn-glycero-3-phosphatidylcholine/ distearoyl-sn-glycero-3-phosphatidylcholine lipid mixture as studied by Fourier transform infrared spectroscopy and differential scanning calorimetry. Biophys J. 1996 Mar;70(3):1373–1379. doi: 10.1016/S0006-3495(96)79695-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Caffrey M., Feigenson G. W. Fluorescence quenching in model membranes. 3. Relationship between calcium adenosinetriphosphatase enzyme activity and the affinity of the protein for phosphatidylcholines with different acyl chain characteristics. Biochemistry. 1981 Mar 31;20(7):1949–1961. doi: 10.1021/bi00510a034. [DOI] [PubMed] [Google Scholar]
  7. Cornea R. L., Thomas D. D. Effects of membrane thickness on the molecular dynamics and enzymatic activity of reconstituted Ca-ATPase. Biochemistry. 1994 Mar 15;33(10):2912–2920. doi: 10.1021/bi00176a022. [DOI] [PubMed] [Google Scholar]
  8. Devaux P. F., Seigneuret M. Specificity of lipid-protein interactions as determined by spectroscopic techniques. Biochim Biophys Acta. 1985 Jun 12;822(1):63–125. doi: 10.1016/0304-4157(85)90004-8. [DOI] [PubMed] [Google Scholar]
  9. Eaton B. R., Dennis E. A. Analysis of phospholipase C (Bacillus cereus) action toward mixed micelles of phospholipid and surfactant. Arch Biochem Biophys. 1976 Oct;176(2):604–609. doi: 10.1016/0003-9861(76)90204-6. [DOI] [PubMed] [Google Scholar]
  10. Fraser D. M., Louro S. R., Horváath L. I., Miller K. W., Watts A. A study of the effect of general anesthetics on lipid-protein interactions in acetylcholine receptor enriched membranes from Torpedo nobiliana using nitroxide spin-labels. Biochemistry. 1990 Mar 20;29(11):2664–2669. doi: 10.1021/bi00463a007. [DOI] [PubMed] [Google Scholar]
  11. Gil T, Mikheev LV. Curvature controlled wetting in two dimensions. Phys Rev E Stat Phys Plasmas Fluids Relat Interdiscip Topics. 1995 Jul;52(1):772–780. doi: 10.1103/physreve.52.772. [DOI] [PubMed] [Google Scholar]
  12. Henderson R., Baldwin J. M., Ceska T. A., Zemlin F., Beckmann E., Downing K. H. Model for the structure of bacteriorhodopsin based on high-resolution electron cryo-microscopy. J Mol Biol. 1990 Jun 20;213(4):899–929. doi: 10.1016/S0022-2836(05)80271-2. [DOI] [PubMed] [Google Scholar]
  13. Henderson R., Unwin P. N. Three-dimensional model of purple membrane obtained by electron microscopy. Nature. 1975 Sep 4;257(5521):28–32. doi: 10.1038/257028a0. [DOI] [PubMed] [Google Scholar]
  14. Horowitz A. D. Exclusion of SP-C, but not SP-B, by gel phase palmitoyl lipids. Chem Phys Lipids. 1995 May 22;76(1):27–39. doi: 10.1016/0009-3084(94)02426-6. [DOI] [PubMed] [Google Scholar]
  15. Hønger T., Jørgensen K., Biltonen R. L., Mouritsen O. G. Systematic relationship between phospholipase A2 activity and dynamic lipid bilayer microheterogeneity. Biochemistry. 1996 Jul 16;35(28):9003–9006. doi: 10.1021/bi960866a. [DOI] [PubMed] [Google Scholar]
  16. Ipsen J. H., Jørgensen K., Mouritsen O. G. Density fluctuations in saturated phospholipid bilayers increase as the acyl-chain length decreases. Biophys J. 1990 Nov;58(5):1099–1107. doi: 10.1016/S0006-3495(90)82452-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Johannsson A., Keightley C. A., Smith G. A., Richards C. D., Hesketh T. R., Metcalfe J. C. The effect of bilayer thickness and n-alkanes on the activity of the (Ca2+ + Mg2+)-dependent ATPase of sarcoplasmic reticulum. J Biol Chem. 1981 Feb 25;256(4):1643–1650. [PubMed] [Google Scholar]
  18. Johannsson A., Smith G. A., Metcalfe J. C. The effect of bilayer thickness on the activity of (Na+ + K+)-ATPase. Biochim Biophys Acta. 1981 Mar 6;641(2):416–421. doi: 10.1016/0005-2736(81)90498-3. [DOI] [PubMed] [Google Scholar]
  19. Jørgensen K., Ipsen J. H., Mouritsen O. G., Bennett D., Zuckermann M. J. The effects of density fluctuations on the partitioning of foreign molecules into lipid bilayers: application to anaesthetics and insecticides. Biochim Biophys Acta. 1991 Aug 26;1067(2):241–253. doi: 10.1016/0005-2736(91)90050-i. [DOI] [PubMed] [Google Scholar]
  20. Jørgensen K., Mouritsen O. G. Phase separation dynamics and lateral organization of two-component lipid membranes. Biophys J. 1995 Sep;69(3):942–954. doi: 10.1016/S0006-3495(95)79968-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jørgensen K., Sperotto M. M., Mouritsen O. G., Ipsen J. H., Zuckermann M. J. Phase equilibria and local structure in binary lipid bilayers. Biochim Biophys Acta. 1993 Oct 10;1152(1):135–145. doi: 10.1016/0005-2736(93)90240-z. [DOI] [PubMed] [Google Scholar]
  22. Kaprelyants A. S. Dynamic spatial distribution of proteins in the cell. Trends Biochem Sci. 1988 Feb;13(2):43–46. doi: 10.1016/0968-0004(88)90024-2. [DOI] [PubMed] [Google Scholar]
  23. Kinnunen P. K. On the principles of functional ordering in biological membranes. Chem Phys Lipids. 1991 Mar;57(2-3):375–399. doi: 10.1016/0009-3084(91)90087-r. [DOI] [PubMed] [Google Scholar]
  24. Knoll W., Ibel K., Sackmann E. Small-angle neutron scattering study of lipid phase diagrams by the contrast variation method. Biochemistry. 1981 Oct 27;20(22):6379–6383. doi: 10.1021/bi00525a015. [DOI] [PubMed] [Google Scholar]
  25. Kusumi A., Hyde J. S. Spin-label saturation-transfer electron spin resonance detection of transient association of rhodopsin in reconstituted membranes. Biochemistry. 1982 Nov 9;21(23):5978–5983. doi: 10.1021/bi00266a039. [DOI] [PubMed] [Google Scholar]
  26. Lehtonen J. Y., Holopainen J. M., Kinnunen P. K. Evidence for the formation of microdomains in liquid crystalline large unilamellar vesicles caused by hydrophobic mismatch of the constituent phospholipids. Biophys J. 1996 Apr;70(4):1753–1760. doi: 10.1016/S0006-3495(96)79738-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lehtonen J. Y., Kinnunen P. K. Phospholipase A2 as a mechanosensor. Biophys J. 1995 May;68(5):1888–1894. doi: 10.1016/S0006-3495(95)80366-8. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. 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]
  29. Mazères S., Schram V., Tocanne J. F., Lopez A. 7-nitrobenz-2-oxa-1,3-diazole-4-yl-labeled phospholipids in lipid membranes: differences in fluorescence behavior. Biophys J. 1996 Jul;71(1):327–335. doi: 10.1016/S0006-3495(96)79228-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Montecucco C., Smith G. A., Dabbeni-sala F., Johannsson A., Galante Y. M., Bisson R. Bilayer thickness and enzymatic activity in the mitochondrial cytochrome c oxidase and ATPase complex. FEBS Lett. 1982 Jul 19;144(1):145–148. doi: 10.1016/0014-5793(82)80588-7. [DOI] [PubMed] [Google Scholar]
  31. Monti J. A., Christian S. T., Shaw W. A. Synthesis and properties of a highly fluorescent derivative of phosphatidylethanolamine. J Lipid Res. 1978 Feb;19(2):222–228. [PubMed] [Google Scholar]
  32. Mouritsen O. G., Bloom M. Mattress model of lipid-protein interactions in membranes. Biophys J. 1984 Aug;46(2):141–153. doi: 10.1016/S0006-3495(84)84007-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Mouritsen O. G., Bloom M. Models of lipid-protein interactions in membranes. Annu Rev Biophys Biomol Struct. 1993;22:145–171. doi: 10.1146/annurev.bb.22.060193.001045. [DOI] [PubMed] [Google Scholar]
  34. Mouritsen O. G., Dammann B., Fogedby H. C., Ipsen J. H., Jeppesen C., Jørgensen K., Risbo J., Sabra M. C., Sperotto M. M., Zuckermann M. J. The computer as a laboratory for the physical chemistry of membranes. Biophys Chem. 1995 Jun-Jul;55(1-2):55–68. doi: 10.1016/0301-4622(94)00142-7. [DOI] [PubMed] [Google Scholar]
  35. Mouritsen O. G., Jørgensen K. Dynamical order and disorder in lipid bilayers. Chem Phys Lipids. 1994 Sep 6;73(1-2):3–25. doi: 10.1016/0009-3084(94)90171-6. [DOI] [PubMed] [Google Scholar]
  36. Muderhwa J. M., Brockman H. L. Lateral lipid distribution is a major regulator of lipase activity. Implications for lipid-mediated signal transduction. J Biol Chem. 1992 Dec 5;267(34):24184–24192. [PubMed] [Google Scholar]
  37. Mustonen P., Virtanen J. A., Somerharju P. J., Kinnunen P. K. Binding of cytochrome c to liposomes as revealed by the quenching of fluorescence from pyrene-labeled phospholipids. Biochemistry. 1987 Jun 2;26(11):2991–2997. doi: 10.1021/bi00385a006. [DOI] [PubMed] [Google Scholar]
  38. Nagle J. F. Area/lipid of bilayers from NMR. Biophys J. 1993 May;64(5):1476–1481. doi: 10.1016/S0006-3495(93)81514-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Oesterhelt D., Stoeckenius W. Isolation of the cell membrane of Halobacterium halobium and its fractionation into red and purple membrane. Methods Enzymol. 1974;31:667–678. doi: 10.1016/0076-6879(74)31072-5. [DOI] [PubMed] [Google Scholar]
  40. Pedersen S., Jørgensen K., Baekmark T. R., Mouritsen O. G. Indirect evidence for lipid-domain formation in the transition region of phospholipid bilayers by two-probe fluorescence energy transfer. Biophys J. 1996 Aug;71(2):554–560. doi: 10.1016/S0006-3495(96)79279-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  42. Piknová B., Pérochon E., Tocanne J. F. Hydrophobic mismatch and long-range protein/lipid interactions in bacteriorhodopsin/phosphatidylcholine vesicles. Eur J Biochem. 1993 Dec 1;218(2):385–396. doi: 10.1111/j.1432-1033.1993.tb18388.x. [DOI] [PubMed] [Google Scholar]
  43. Pink D. A., Green T. J., Chapman D. Raman scattering in bilayers of saturated phosphatidylcholines. Experiment and theory. Biochemistry. 1980 Jan 22;19(2):349–356. doi: 10.1021/bi00543a016. [DOI] [PubMed] [Google Scholar]
  44. Rehorek M., Heyn M. P. Binding of all-trans-retinal to the purple membrane. Evidence for cooperativity and determination of the extinction coefficient. Biochemistry. 1979 Oct 30;18(22):4977–4983. doi: 10.1021/bi00589a027. [DOI] [PubMed] [Google Scholar]
  45. Rigaud J. L., Pitard B., Levy D. Reconstitution of membrane proteins into liposomes: application to energy-transducing membrane proteins. Biochim Biophys Acta. 1995 Oct 10;1231(3):223–246. doi: 10.1016/0005-2728(95)00091-v. [DOI] [PubMed] [Google Scholar]
  46. Sankaram M. B., Marsh D., Thompson T. E. Determination of fluid and gel domain sizes in two-component, two-phase lipid bilayers. An electron spin resonance spin label study. Biophys J. 1992 Aug;63(2):340–349. doi: 10.1016/S0006-3495(92)81619-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Sankaram M. B., Thompson T. E. Deuterium magnetic resonance study of phase equilibria and membrane thickness in binary phospholipid mixed bilayers. Biochemistry. 1992 Sep 8;31(35):8258–8268. doi: 10.1021/bi00150a020. [DOI] [PubMed] [Google Scholar]
  48. Schram V., Thompson T. E. Influence of the intrinsic membrane protein bacteriorhodopsin on gel-phase domain topology in two-component phase-separated bilayers. Biophys J. 1997 May;72(5):2217–2225. doi: 10.1016/S0006-3495(97)78865-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Schram V., Tocanne J. F., Lopez A. Influence of obstacles on lipid lateral diffusion: computer simulation of FRAP experiments and application to proteoliposomes and biomembranes. Eur Biophys J. 1994;23(5):337–348. doi: 10.1007/BF00188657. [DOI] [PubMed] [Google Scholar]
  50. Sperotto M. M., Mouritsen O. G. Lipid enrichment and selectivity of integral membrane proteins in two-component lipid bilayers. Eur Biophys J. 1993;22(5):323–328. doi: 10.1007/BF00213555. [DOI] [PubMed] [Google Scholar]
  51. Sperotto M. M., Mouritsen O. G. Mean-field and Monte Carlo simulation studies of the lateral distribution of proteins in membranes. Eur Biophys J. 1991;19(4):157–168. doi: 10.1007/BF00196342. [DOI] [PubMed] [Google Scholar]
  52. Sperotto M. M., Mouritsen O. G. Monte Carlo simulation studies of lipid order parameter profiles near integral membrane proteins. Biophys J. 1991 Feb;59(2):261–270. doi: 10.1016/S0006-3495(91)82219-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Tocanne J. F., Cézanne L., Lopez A., Piknova B., Schram V., Tournier J. F., Welby M. Lipid domains and lipid/protein interactions in biological membranes. Chem Phys Lipids. 1994 Sep 6;73(1-2):139–158. doi: 10.1016/0009-3084(94)90179-1. [DOI] [PubMed] [Google Scholar]
  54. Tu K., Tobias D. J., Blasie J. K., Klein M. L. Molecular dynamics investigation of the structure of a fully hydrated gel-phase dipalmitoylphosphatidylcholine bilayer. Biophys J. 1996 Feb;70(2):595–608. doi: 10.1016/S0006-3495(96)79623-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Yguerabide J. Theory for establishing proximity relations in biological membranes by excitation energy transfer measurements. Biophys J. 1994 Mar;66(3 Pt 1):683–693. doi: 10.1016/s0006-3495(94)80842-2. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES