Sensing isothermal changes in the lateral pressure in model membranes using di-pyrenyl phosphatidylcholine

Abstract

In this work we present data from a homologous series of di-pyrenyl phosphatidylcholine (dipyPC) probes which can sense lateral pressure variations in the chain region of the amphiphilic membrane (lateral pressures are tangential to the interface). The dipyPC has pyrene moieties attached to the ends of equal length acyl chains on a phosphatidylcholine molecule. Ultraviolet stimulation produces both monomer and excimer fluorescence from pyrene. At low dilutions of dipyPC in model membranes the excimer signal is entirely intra-molecular and since it depends on the frequency with which the pyrene moieties are brought into close proximity, the relative intensity of the excimer to monomer signal, η, is a measure of the pressure. We synthesised or purchased dipyPC probes with the pyrene moieties attached to acyl chains having 4, 6, 8 and 10 carbon atoms and then measured η in fully hydrated bilayers composed of dioleoylphosphatidylcholine and dioleoylphosphatidylethanolamine (DOPC and DOPE respectively). Although the resolution of our measurements of lateral pressure as a function of distance into the monolayer was limited, we did observe a dip in the excimer signal in the region of the DOPC/DOPE cis double bond. As we isothermally increased the DOPE composition, and hence the desire for interfacial curvature, we observed, as expected, that the net excimer signal increased. However this net increase was apparently brought about by a transfer of pressure from the region around the glycerol backbone to the region near the chain ends, with the lateral pressure dropping above the cis double bond but increasing at a greater rate beyond the double bond.

References

1. W. Helfrich, Z. Naturforsch., 1973, 28c, 693.
2. S. M. Gruner, Proc. Natl. Acad. Sci. USA, 1985, 82, 3665.
3. J. M. Seddon, Biochim. Biophys. Acta, 1990, 1031, 1.
4. G. Lindblom, A. Wieslander, M. Sjoelund, G. Wikander and A. Wieslander, Biochemistry, 1986, 25, 7502.
5. S. L. Keller, S. M. Bezrukov, S. M. Gruner, M. W. Tate, I. Vodyanoy and V. A. Parsegian, Biophys. J., 1993, 65, 23.
6. R. M. Epand, Chem. Phys. Lipids, 1996, 81, 101.
7. P. J. Booth, M. L. Riley, S. L. Flitsch, R. H. Templer, A. Farooq, A. R. Curran, N. Chadborn and P. Wright, Biochemistry, 1997, 36, 197.
8. G. S. Attard, R. H. Templer, W. S. Smith, A. N. Hunt and S. Jackowski, Nature, 1999, submitted; G. S. Attard, W. S. Smith, R. H. Templer, A. N. Hunt and S. Jackowski, Biochem. Soc. Trans., 1998, 26, 5230.
9. W. Helfrich, in Physics of defects, ed. R. Balian, M. Kléman and J. P. Poirier, North-Holland, Amsterdam, 1981, p. 715.
10. I. Szleifer, A. Benshaul and W. M. Gelbart, J. Phys. Chem., 1990, 94, 5081.
11. I. Szleifer, D. Kramer, A. Benshaul, W. M. Gelbart and S. A. Safran, J. Chem. Phys., 1990, 92, 6800.
12. I. Szleifer, D. Kramer, A. Benshaul, D. Roux and W. M. Gelbart, Phys. Rev. Lett., 1988, 60, 1966.
13. S. M. Gruner, M. W. Tate, G. L. Kirk, P. T. C. So, D. C. Turner, D. T. Keane, C. P. S. Tilcock and P. R. Cullis, Biochemistry, 1988, 27, 2853.
14. J. P. Birks, Photophysics of aromatic molecules, Wiley, London, 1970.
15. J. B. Birks and L. G. Christophorou, Proc. R. Soc. London, Ser. A, 1963, 274, 552.
16. J. B. Birks, D. J. Dyson and I. H. Munro, Proc. R. Soc. London, Ser. A, 1963, 275, 575.
17. J. B. Birks and L. G. Christophorou, Proc. R. Soc. London, Ser. A, 1964, 277, 571.
18. J. B. Birks, D. J. Dyson and T. A. King, Proc. R. Soc. London, Ser. A, 1964, 277, 270.
19. J. B. Birks, M. D. Lumb and I. H. Munro, Proc. R. Soc. London, Ser. A, 1964, 280, 289.
20. K. H. Cheng, L. Ruymgaart, L.-I. Liu, P. Somerharju and I. P. Sugár, Biophys. J., 1994, 67, 914.
21. J. Sunamoto, H. Kondo, T. Nomura and H. Okamoto, J. Am. Chem. Soc., 1980, 102, 1146.
22. J. Sunamoto, T. Nomura and H. Okamoto, Bull. Chem. Soc. Jpn., 1980, 53, 2768.
23. K. H. Cheng, S.-Y. Chen, P. Butko, B. W. Van Der Meer and P. Somerharju, Biophys. Chem., 1991, 39, 137.
24. P. Butko and K. H. Cheng, Chem. Phys. Lipids, 1992, 62, 39.
25. M. Sassaroli, M. Vaukhonen, P. Somerharju and S. Scarlata, Biophys. J., 1993, 64, 137.
26. S.-Y. Chen, K. H. Cheng and B. W. Van Der Meer, Biochemistry, 1992, 31, 3759.
27. Z. Chen and R. P. Rand, Biophys. J., 1997, 73, 267.
28. M. Vaukhonen and P. Somerharju, Chem. Phys. Lipids, 1990, 52, 207.
29. H. S. Henderson and P. N. Rauk, Anal. Biochem., 1981, 116, 553.
30. G. Rumbles, unpublished data.
31. J. Y. A. Lehtonen and P. K. J. Kinnunen, Biophys. J., 1981, 66, 1981.
32. K. H. Madan, MSc thesis, Imperial College, London University, 1991.
33. R. P. Rand, N. L. Fuller, S. M. Gruner and V. A. Parsegian, Biochemistry, 1990, 29, 76.
34. A. Ben-Shaul, personal communication.
35. F. Reiss-Husson and V. Luzzati, J. Phys. Chem., 1964, 68, 3504.
36. V. Luzzati and F. Husson, J. Cell. Biol., 1962, 12, 207.