Abstract
Cholesterol is a key player in regulating physico-chemical properties of cellular membranes and, thereby, ensuring cell viability. In particular, lipid-cholesterol interactions may provide important information on the spatio-temporal organization of membrane components. Here, we apply confocal imaging and Fluorescence Correlation Spectroscopy (FCS) to Giant Unilamellar Vesicles (GUVs) composed of binary mixtures of lipids and cholesterol.
The effect of cholesterol on lipid dynamics and molecular packing order of unsaturated, monounsaturated, fully saturated (with both low and high phase transition temperatures, Tm) glycero-phospholipids and sphingomyelin has been investigated. We show that, for unsaturated glycerophospholipids, the decrease of the lipid diffusion coefficient as a result of the interaction with cholesterol does not depend on the fatty acid chain length. However, the values of the diffusion coefficient change as a function of chain length. The monounsaturated phospholipid palmitoyl-oleoyl-phosphatidylcholine (POPC) exhibits a dynamic behavior very similar to the unsaturated dioleoyl-phosphatidylcholine (DOPC). By contrast, for saturated (low Tm) glycero-phospholipids, cholesterol causes a decrease of lipid mobility in a chain length-dependent manner.
FCS can be employed as a valuable tool to study lipid-sterol interactions and their effect on lipid dynamics, molecular packing and degree of conformational order.
Similar content being viewed by others
References
de Kruijff B et al (1985) Lipid polymorphism and membrane function. The enzymes of biological membranes, vol 1, In: Martonosi AN (ed) Membrane structure and dynamics, 2nd edn. Plenum Press, New York, pp 131–204
Yeagle PL (1985) Cholesterol and the cell membrane. Biochim Biophys Acta 822:267–287
Bretscher MS, Munro S (1993) Cholesterol and the Golgi apparatus. Science 261:1280–1281
Simons K, van Meer G (1988) Lipid sorting in epithelial cells. Biochemistry 27:6197–6202
Simons K, Ikonen E (1997) Functional rafts in cell membranes. Nature 387:569–572
Simons K, Ikonen E (2000) How cells handle cholesterol. Science 290:1721–1726
Veatch SL, Keller SL (2005) Seeing spots: complex phase behavior in simple membranes. Biochim et Biophys Acta (in press)
Angelova MI, Dimitrov DS (1986) Liposome electroformation, Faraday Discuss. Chem Soc 81:303–308
Menger FM, Keiper JS (1998) Chemistry and physics of giant vesicles as biomembrane models. Curr Op Chem Biol 2:726– 732
Dietrich C, Bagatolli LA, Volovyk ZN, Thompson NL, Levi M, Jacobson K, Gratton E (2001) Lipid rafts reconstituted in model membranes. Biophys J 80:1417–1428
Samsonov AV, Mihalyov I, Cohen FS (2001) Characterization of cholesterol-sphingomyeliindomains and their dynamicsin bilayer membranes. Biophys J 81:1486–1500
Kahya N, Scherfeld D, Bacia K, Poolman B, Schwille P (2003) Probing lipid mobility of raft-exhibiting model membranes by Fluorescence Correlation Spectroscopy. J Biol Chem 278:28109–28115
Rigler R, Elson E (eds) (2001) Fluorescence Correlation Spectroscopy: theory and applications. Springer, Berlin
Schwille P (2001) Fluorescence Correlation Spsctroscopy and its potential for intracellular applications. Cell Biochem Biophys 34:383–408
Scherfeld D, Kahya N, Schwille P (2003) Lipid dynamics and domain formation in model membranes composed of ternary mixtures of saturated and unsaturated phosphatidylcholines and cholesterol. Biophys J 85:3758–3768
Benda A, Beneš M, Mareček V, Lhotsky A, Th Hermens W, Hof M (2003) How to determine diffusion coefficients in planar phospholipid systems by confocal fluorescence correlation spectroscopy. Langmuir 19:4120–4126
Magde D, Elson EL, Webb WW (1972) Thermodynamic fluctuations in a reacting system-measurement by fluorescence correlation spectroscopy. Phys Rev Lett 29:705–708
Colladom MI, Goni FM, Alonso A, Marsh D (2005) Domain formation in sphingomyelin/cholesterol mixed membranes studied by spin-label electron spin resonance spectroscopy. Biochemistry 44:4911–4918
Loura LMS, Fedorov A, Prieto M (2001) Fluid-fluid membrane microheterogeneity: a fluorescence resonance energy transfer study. Biophys J 80:776–788
Feigenson GW, Buboltz JT (2001) Ternary phase diagram of dipalmitoyl-PC/dilauroyl-PC/cholesterol: nanoscopic domain formation driven by cholesterol. Biohys J 80:2775– 2788
Vist MR, Davis JH (1990) Phase equilibria of cholesterol/dipalmitoylphosphatidylcholine mixtures: 2H nuclear magnetic resonance and differential scanning calorimetry. Biochemistry 29:451–464
Veatch SL, Keller SL (2003) Separation of liquid phases in giant vesicles of ternary mixtures of phospholipids and cholesterol. Biophys J 85:3074–3083
Korlach J, Schwille P, Webb WW, Feigenson GW (1999) Characterization of lipid bilayer phases by confocal microscopy and fluorescence correlation spectroscopy. Proc Natl Acad Sci USA 96:8461–8466
Kahya N, Scherfeld D, Bacia K, Schwille P (2003) Lipid domain formation and dynamics in giant unilamellar vesicles explored by fluorescence correlation spectroscopy. J Struct Biol 147:77– 89
Schütz GJ, Schindler H, Schmidt T (1997) Single-molecule microscopy on model membranes reveals anomalous diffusion. Biophys J 73:1073–1080
Filippov A, Oradd G, Lindblom G (2004) Lipid lateral diffusion in ordered and disordered phases in raft mixtures. Biophys J 86:891–896
Feingold L (1993) Cholesterol in membrane models. CRC, Ann Arbor, MI
Wu W-G, Chi L-M (1991) Conformational change of cholesterol side chain in lipid bilayers. J Am Chem Soc 113:4683–4685
Brzustowicz MR, Cherezov V, Zerouga M, Caffrey M, Stillwell W, Wassall SR (2002) Controlling membrane cholesterol content. A role for polyunsaturated (docosahexaenoate) phospholipids. Biochemistry 41:12509–12519
de Kruijff B, Demel RA, Slotboom AJ, van Deenen LLM, Rosenthal AF (1973) The effect of the polar headgroup on the lipid-cholesterol interaction: A monolayer and differential scanning calorimetry study. Biochim Biophys Acta 307:1–19
Veatch SL, Keller SL (2005) Miscibility of phase diagrams of giant vesicles containing sphingomyelin. Phys Rev Lett 94:148101-1–148101-4
de Almeida RFM, Fedorov A, Prieto M (2003) Sphingomyelin/phosphatidylcholine/cholesterol phase diagram: boundaries and composition of lipid rafts. Biophys J 85:2406–2416
de Almeida RFM, Loura LMS, Fedorov A, Prieto M (2005) Lipid rafts have different sizes depending on membrane composition: a time-resolved fluorescence resonance energy transfer study. J Mol Biol 346:1109–1120
McMullen TPW, McElhaney RN (1995) New aspects of the interaction of cholesterol with dipalmitoylphosphatidylcholine bilayers as revealed by high-sensitivity differential scanning calorimetry. Biochim Biophys Acta 1234:90–98
Ohvo-Rekilä HB, Ramstedt B, Leppimäki P, Slotte JP (2002) Cholesterol interactions with phospholipids in membranes. Progr Lip Res 41:66–97
Acknowledgments
We thank Dick Hoekstra, Lucie Kalvodova, and Dennis Merkle for stimulating discussions.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Kahya, N., Schwille, P. How Phospholipid-Cholesterol Interactions Modulate Lipid Lateral Diffusion, as Revealed by Fluorescence Correlation Spectroscopy. J Fluoresc 16, 671–678 (2006). https://doi.org/10.1007/s10895-006-0108-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10895-006-0108-6