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Lateral diffusion of erythrocyte phospholipids in model membranes comparison between inner and outer leaflet components

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Abstract

The physical properties of lipid bilayers with a similar composition to the outer and inner leaflets of the human erythrocyte membrane have been examined in protein-free model systems. The outer leaflet (OL) was represented by a phospholipid mixture containing phosphatidylcholine and sphingomyelin extracted from human erythrocytes, while a mixture of phosphatidylcholine, phosphatidylserine and phosphatidylethanolamine represented the inner leaflet (IL). The ratio of cholesterol to phospholipid was varied in both mixtures. The lateral diffusion coefficient of fluorescent phospholipids diluted in such lipid mixtures was determined by the modulated fringe pattern photobleaching technique. Contrast curves with a single exponential decay, indicative of homogeneous samples, were obtained only for temperatures above 15 °C and for a cholesterol to phospholipid molar ratio below 0.8. The rate of lateral diffusion was approximately five times faster in IL than in OL multilayers, in agreement with former results obtained in human erythrocytes (Morrot et al. 1986). Varying the cholesterol to phospholipid ratio from 0 to 0.8 (mol/mol) enabled us to decrease the diffusion constant by only a factor of approximately 2 for both IL and OL mixtures. The order parameter of a spin-labeled phospholipid was determined in the different systems and found to be systematically smaller in IL mixtures than in OL mixtures. The present study indicates that the difference in lipid diffusivity of the two erythrocyte leaflets may be accounted for solely by a difference in phospholipid composition, and may be independent of cholesterol and protein asymmetry.

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Abbreviations

OL:

outer leaflet

IL:

inner leaflet

RBC:

red blood cell

NBD-PC:

1-acyl-2-[12-(7-nitrobenz-2-oxa-1,3-diazol-4-yl) amino] dodecanoyl phosphatidylcholine

NBD-PE:

1-acyl-2-[12-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] dodecanoyl phosphatidylethanolamine

NBD-PS:

1-acyl-2-[12-(7-nitrobenz-2-oxy-1,3-diazol-4-yl)amino] dodecanoyl phosphatidylserine

DMPC:

1,2 dimyristoyl-sn-glycero-3-phosphocholine

DMPS:

1,2 dimyristoyl-snglycero-3-phosphoserine

PC:

phosphatidyleholine

C/P:

cholesterol over phospholipid molar ratio

D:

lateral diffusion coefficient

S:

order parameter

ESR:

electron spin resonance

NMR:

nuclear magnetic resonance

EDTA:

ethylene diamine tetraacetic acid

TRIS:

tris-(hydroxymethyl)amino ethane

References

  • Bitbol M, Dempsey C, Watts A, Devaux PF (1989) Weak interaction of spectrin with phosphatidylcholine-phosphatidylserine multilayer: a 2H and 31P NMR study. FEBS Lett 244:217–222

    Google Scholar 

  • Calvez JY, Zachowski A, Herrmann A, Morrot G, Devaux PF (1988) Asymmetric distribution of phospholipids in spectrin poor erythrocyte vesicles. Biochemistry 27:5666–5670

    Google Scholar 

  • Chang CH, Takeuchi H, Ito T, Machida K, Ohnishi S-I (1981) Lateral mobility of erythrocyte membrane proteins studied by the fluorescence photobleaching recovery technique. J Biochim 90:997–1004

    Google Scholar 

  • Clegg RM, Vaz WLC (1985) Translational diffusion of proteins and lipids in artificial lipid bilayer membranes. A comparison of experiment with theory. In: Watts A, De Pont JJHHM (eds) Progress in protein-lipid interactions, vol 1. Elsevier, Amsterdam, pp 173–229

    Google Scholar 

  • Cogan U, Schachter D (1981) Asymmetry of lipid dynamics in human erythrocyte membranes studied with impermeant fluorophores. Biochemistry 20:6396–6403

    Google Scholar 

  • Comfurius P, Zwaal RFA (1977) The enzymatic synthesis of phosphatidylserine and purification by CM-cellulose column chromatography. Biochim Biophys Acta 488:36–42

    Google Scholar 

  • Cullis PR, Grathwohl C (1977) Hydrocarbon phase transitions and lipid-protein interactions in the erythrocyte membrane. A 31P and fluorescence study. Biochim Biophys Acta 471:213–226

    Google Scholar 

  • Davoust J, Devaux PF, Léger L (1982) Fringe pattern photobleaching, a new method for the measurement of transport coefficients of biological macromolecules. EMBO J 1:1233–1238

    Google Scholar 

  • Derzko Z, Jacobson K (1980) Comparative lateral diffusion of fluorescent lipid analogues in phospholipid multibilayers. Biochemistry 19:6050–6057

    Google Scholar 

  • Devaux PF (1988) Phospholipid flippases. FEBS Lett 234:8–12

    Google Scholar 

  • Devaux PF, McConnell HJM (1972) Lateral diffusion in spin labeled phosphatidylcholine multilayers. J Am Chem Soc 94: 4475–4481

    Google Scholar 

  • Finean JB (1969) Biophysical contributions to membrane structure. Q Rev Biophys 2:1–23

    Google Scholar 

  • Gaffney BJ (1976) Practical considerations for the calculation of order parameters for fatty acid or phospholipid spin labels in membranes. In: Berliner LJ (ed) Spin labelling, theory and applications. Academic Press, New York, pp 567–571

    Google Scholar 

  • Galla HJ, Hartmann W, Theilen U, Sackmann E (1979) On two dimensional passive random walk in lipid bilayers and fluid pathways in biomembranes. J Membr Biol 48:215–236

    Google Scholar 

  • Golan DE, Alecio MR, Veatch WR, Rando RR (1984) Lateral mobility of phospholipid and cholesterol in the human erythrocyte membrane: Effects of protein-lipid interactions. Biochemistry 23:332–339

    Google Scholar 

  • Haest CWM (1982) Interactions between membrane skeleton proteins and the intrinsic domain of the erythrocyte membrane. Biochim Biophys Acta 694:331–352

    Google Scholar 

  • Hale JE and Schroeder F (1982) Asymmetric transbilayer distribution of sterol across plasma membranes determined by fluorescence quenching of dehydroergosterol. Eur J Biochem 122: 649–661

    Google Scholar 

  • Henis YI, Rimon G, Felder S (1982) Lateral mobility of phospholipids in turkey erythrocytes Implication for adenylate cyclase activation. J Biol Chem 257:1407–1411

    Google Scholar 

  • Hope MJ, Cullis PR (1979) The bilayer stability of inner monolayer lipids from the human erythrocyte. FEBS Lett 107:323–326

    Google Scholar 

  • Hubbell WL, McConnell HM (1971) Molecular motion in spin-labeled phospholipids and membranes. J Am Chem Soc 93: 314–326

    Google Scholar 

  • Kuo AL, Wade CG (1979) Lipid lateral diffusion by pulsed nuclear magnetic resonance. Biochemistry 18:2300–2308

    Google Scholar 

  • Lindblom G, Johansson LBA, Arvidson G (1981) Effect of cholesterol in membranes pulsed nuclear magnetic resonance measurements of lipid lateral diffusion. Biochemistry 20: 2204–2207

    Google Scholar 

  • Maksymiw R, Sui S-F, Gaub H, Sackmann E (1987) Electrostatic coupling of spectrin dimers to phosphatidylserine containing lipid lamellae. Biochemistry 26:2990–2993

    Google Scholar 

  • McCown JT, Evans E, Diehl S, Wiles HC (1981) Degree of hydration and lateral diffusion in phospholipid multibilayers. Biochemistry 20:3134–3138

    Google Scholar 

  • Morrot G, Cribier S, Devaux, PF, Geldwerth D, Davoust J, Bureau JF, Fellmann P, Herve P, Frilley B (1986) Asymmetric lateral mobility of phospholipids in the human erythrocyte membrane. Proc Natl Acad Sci USA 83:6863–6867

    Google Scholar 

  • Morrot G, Hervé P, Zachowski A, Fellmann P, Devaux PF (1989) Aminophospholipid translocase of human erythrocytes: phospholipid substrate specificity and effect of cholesterol. Biochemistry 28:3456–3462

    Google Scholar 

  • Rose HG, Oklander M (1965) Improved procedure for the extraction of lipids from human erythrocytes. J Lipid Res 6:428–431

    Google Scholar 

  • Roux M, Neumann J-M, Bloom M, Devaux PF (1988) 2H and 31p NMR study of pentalysine interaction with headgroup deuterated phosphatidylcholine and phosphatidylserine. Eur J Biophys 16:267–273

    Google Scholar 

  • Rubenstein JLR, Smith BA, McConnell HM (1979) Lateral diffusion in binary mixtures of cholesterol and phosphatidylcholines. Proc Natl Acad Sci USA 76:15–18

    Google Scholar 

  • Saffman PG, Delbrück M (1975) Brownian motion in biological membranes. Proc Natl Acad Sci USA 72:3111–3113

    Google Scholar 

  • Schachter D, Cogan U, Abbot RE (1982) Asymmetry of lipid dynamics in human erythrocyte membranes studied with permeant fluorophores. Biochemistry 21:2146–2150

    Google Scholar 

  • Schlegel RA, Williamson P (1988) Role of phospholipid asymmetry in cellular membrane fusion. In: Ohki S, Doyle D, Flanagan TD, Hui SW, Mayhew E (eds) Molecular mechanisms of membrane fusion. Plenum Press, New York, pp 289–301

    Google Scholar 

  • Schlegel RA, Phelps BM, Waggoner A, Terada L, Williamson P (1980) Binding of merocyanine 540 to normal and leukemic erythroid cells. Cells 20:321–328

    Google Scholar 

  • Schroeder F (1981) Use of fluorescent sterol to probe the transbilayer distribution of sterols in biological membranes. FEBS Lett 135:127–130

    Google Scholar 

  • Schroeder F, Nemecz G (1989) Transmembrane cholesterol distribution. In: Esfahani M, Swaney J (eds) Advances in cholesterol metabolism (in press)

  • Seelig J, Borle F, Cross TA (1985) Magnetic ordering of phospholipid membranes. Biochim Biophys Acta 814:195–198

    Google Scholar 

  • Seigneuret M, Devaux PF (1984) ATP-dependant asymmetric distribution of spin labeled phospholipids in the erythrocyte membrane. Relation to shape changes. Proc Natl Acad Sci USA 81:3751–3755

    Google Scholar 

  • Seigneuret M, Zachowski A, Hermann A, Devaux PF (1984) Asymmetric lipid fluidity in human erythrocyte membrane: new spin-label evidence. Biochemistry 23:4271–4275

    Google Scholar 

  • Singleton WS, Gray MS, Brown ML, White JL (1965) Chromatographically homogeneous lecithin from egg phospholipids. J Am Oil Chem Soc 42:53–56

    Google Scholar 

  • Skin Y-K, Freed JH (1989) Dynamic imaging of lateral diffusion by electron spin resonance and study of rotational dynamics in model membranes. Effect of cholesterol. Biophys J 55: 537–550

    Google Scholar 

  • Speyer JB, Sripada PK, Das Gupta SK, Shipley GG, Griffin RG (1987) Magnetic orientation of sphingomyelin-lecithin bilayers. Biophys J 51:687–691

    Google Scholar 

  • Tanaka K, Ohnishi S-I (1976) Heterogeneity in the fluidity of intact erythrocyte membrane and its homogenization upon hemolysis. Biochim Biophys Acta 426:218–231

    Google Scholar 

  • Thompson NL, Axelrod D (1980) Reduced lateral mobility of a fluorescent lipid probe in cholesterol-depleted erythrocyte membrane. Biochim Biophys Acta 597:155–165

    Google Scholar 

  • Vaz WLC, Derzko ZI, Jacobson KA (1982) Photobleaching measurements of the lateral diffusion of lipids and proteins in artificial bilayer membranes. In: Poste G, Nicolson G (eds) Membrane reconstitution, Elsevier, Amsterdam, pp. 83–135

    Google Scholar 

  • Vaz WLC, Clegg R, Hallmann D (1985) Transitional diffusion of lipids in liquid crystalline phase phosphatidylcholine multibilayers. A comparison of experiment with theory. Biochemistry 24:781–786

    Google Scholar 

  • Verkleij AJ, Zwaal RFA, Roeloefsen B, Comfurius P, Kastelijn D, van Deenen LLM (1973) The asymmetric distribution of phospholipids in the human red cell membrane. A combined study using phospholipases and freeze-etching electronmicroscopy. Biochim Biophys Acta 323:178–193

    Google Scholar 

  • Ways P, Hanahan DJ (1964) Characterization and quantification of red cell lipids in normal man. J Lipids Res 5:318–328

    Google Scholar 

  • Williamson P, Bateman J, Kozarsky K, Mattocks K, Hermanowicz N, Choe HR, Schlegel RA (1982) Involvement of spectrin in the maintenance of phase-state asymmetry in the erythrocyte membrane. Cell 30:725–733

    Google Scholar 

  • Wu E-S, Jacobson K, Papahadjopoulos D (1977) Lateral diffusion in phospholipid multilayers measured by fluorescence recovery after photobleaching. Biochemistry 16:3936–3941

    Google Scholar 

  • Zachowski A, Devaux PF (1989) Bilayer asymmetry and lipid transport across biomembranes. Comments in molecular and cellular biophys. 6:63–90

    Google Scholar 

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Authors and Affiliations

  1. Institut de Biologie Physico-Chimique, 13 rue Pierre et Marie Curie, F-75005, Paris, France

    S. Cribier, G. Morrot & P. F. Devaux

  2. Departement de Biologie, Service de Biophysique, CEN-Saclay, F-91191, Gif-sur-Yvette, France

    J.-M. Neumann

Authors
  1. S. Cribier
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  2. G. Morrot
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  3. J.-M. Neumann
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  4. P. F. Devaux
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Cribier, S., Morrot, G., Neumann, JM. et al. Lateral diffusion of erythrocyte phospholipids in model membranes comparison between inner and outer leaflet components. Eur Biophys J 18, 33–41 (1990). https://doi.org/10.1007/BF00185418

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  • DOI: https://doi.org/10.1007/BF00185418

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