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. 1984 Jun;81(12):3751–3755. doi: 10.1073/pnas.81.12.3751

ATP-dependent asymmetric distribution of spin-labeled phospholipids in the erythrocyte membrane: relation to shape changes.

M Seigneuret, P F Devaux
PMCID: PMC345297  PMID: 6587389

Abstract

Spin-labeled analogs of phosphatidylcholine, phosphatidylserine, and phosphatidylethanolamine have been used to study phospholipid transverse diffusion and asymmetry in the human erythrocyte membrane. Ascorbate reduction was used to assess the transbilayer distribution of the labels. All three spin-labeled phospholipids initially incorporated into the outer leaflet of the membrane. On fresh erythrocytes at 5 degrees C, the phosphatidylcholine label remained mainly in the outer leaflet. In contrast, the phosphatidylserine and phosphatidylethanolamine labels underwent rapid transverse diffusion that led to their asymmetric distribution in favor of the inner leaflet. The latter effect was reversibly inhibited after ATP depletion of the erythrocytes and could be reproduced on resealed erythrocyte ghosts only if hydrolyzable Mg-ATP was included in the internal medium. It is suggested that an ATP-driven transport of amino phospholipids toward the inner leaflet could be the major cause of the phospholipid asymmetry in the erythrocyte membrane. It is also proposed that the same mechanism could explain the ATP requirement of the maintenance of the erythrocyte membrane discoid shape.

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Selected References

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  1. Anderson J. M., Tyler J. M. State of spectrin phosphorylation does not affect erythrocyte shape or spectrin binding to erythrocyte membranes. J Biol Chem. 1980 Feb 25;255(4):1259–1265. [PubMed] [Google Scholar]
  2. Beck J. S. Relations between membrane monolayers in some red cell shape transformations. J Theor Biol. 1978 Dec 21;75(4):487–501. doi: 10.1016/0022-5193(78)90358-2. [DOI] [PubMed] [Google Scholar]
  3. Bergmann W. L., Dressler V., Haest C. W., Deuticke B. Cross-linking of SH-groups in the erythrocyte membrane enhances transbilayer reorientation of phospholipids. Evidence for a limited access of phospholipids to the reorientation sites. Biochim Biophys Acta. 1984 Jan 25;769(2):390–398. doi: 10.1016/0005-2736(84)90322-5. [DOI] [PubMed] [Google Scholar]
  4. Birchmeier W., Singer S. J. On the mechanism of ATP-induced shape changes in human erythrocyte membranes. II. The role of ATP. J Cell Biol. 1977 Jun;73(3):647–659. doi: 10.1083/jcb.73.3.647. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bretscher M. S. Phosphatidyl-ethanolamine: differential labelling in intact cells and cell ghosts of human erythrocytes by a membrane-impermeable reagent. J Mol Biol. 1972 Nov 28;71(3):523–528. doi: 10.1016/s0022-2836(72)80020-2. [DOI] [PubMed] [Google Scholar]
  6. Comfurius P., Zwaal R. F. The enzymatic synthesis of phosphatidylserine and purification by CM-cellulose column chromatography. Biochim Biophys Acta. 1977 Jul 20;488(1):36–42. doi: 10.1016/0005-2760(77)90120-5. [DOI] [PubMed] [Google Scholar]
  7. Davoust J., Seigneuret M., Hervé P., Devaux P. F. Collisions between nitrogen-14 and nitrogen-15 spin-labels. 1. Lipid-lipid interactions in model membranes. Biochemistry. 1983 Jun 21;22(13):3137–3145. doi: 10.1021/bi00282a016. [DOI] [PubMed] [Google Scholar]
  8. Deuticke B., Poser B., Lütkemeier P., Haest C. W. Formation of aqueous pores in the human erythrocyte membrane after oxidative cross-linking of spectrin by diamide. Biochim Biophys Acta. 1983 Jun 10;731(2):196–210. doi: 10.1016/0005-2736(83)90009-3. [DOI] [PubMed] [Google Scholar]
  9. Franck P. F., Roelofsen B., Op den Kamp J. A. Complete exchange of phosphatidylcholine from intact erythrocytes after protein crosslinking. Biochim Biophys Acta. 1982 Apr 23;687(1):105–108. doi: 10.1016/0005-2736(82)90176-6. [DOI] [PubMed] [Google Scholar]
  10. Féo C. J., Leblond P. F. The discocyte-echinocyte transformation: comparison of normal and ATP-enriched human erythrocytes. Blood. 1974 Nov;44(5):639–647. [PubMed] [Google Scholar]
  11. Gordesky S. E., Marinetti G. V. The asymetric arrangement of phospholipids in the human erythrocyte membrane. Biochem Biophys Res Commun. 1973 Feb 20;50(4):1027–1031. doi: 10.1016/0006-291x(73)91509-x. [DOI] [PubMed] [Google Scholar]
  12. Haest C. W., Deuticke B. Possible relationship between membrane proteins and phospholipid asymmetry in the human erythrocyte membrane. Biochim Biophys Acta. 1976 Jun 17;436(2):353–365. doi: 10.1016/0005-2736(76)90199-1. [DOI] [PubMed] [Google Scholar]
  13. Haest C. W., Plasa G., Kamp D., Deuticke B. Spectrin as a stabilizer of the phospholipid asymmetry in the human erythrocyte membrane. Biochim Biophys Acta. 1978 May 4;509(1):21–32. doi: 10.1016/0005-2736(78)90004-4. [DOI] [PubMed] [Google Scholar]
  14. Hayashi H., Jarrett H. W., Penniston J. T. Peripheral proteins and smooth membrane from erythrocyte ghosts. Segregation of ATP-utilizing enzymes into smooth membrane. J Cell Biol. 1978 Jan;76(1):105–115. doi: 10.1083/jcb.76.1.105. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Johnson R. M., Taylor G., Meyer D. B. Shape and volume changes in erythrocyte ghosts and spectrin-actin networks. J Cell Biol. 1980 Aug;86(2):371–376. doi: 10.1083/jcb.86.2.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kornberg R. D., McConnell H. M. Inside-outside transitions of phospholipids in vesicle membranes. Biochemistry. 1971 Mar 30;10(7):1111–1120. doi: 10.1021/bi00783a003. [DOI] [PubMed] [Google Scholar]
  17. Lange Y., Hadesman R. A., Steck T. L. Role of the reticulum in the stability and shape of the isolated human erythrocyte membrane. J Cell Biol. 1982 Mar;92(3):714–721. doi: 10.1083/jcb.92.3.714. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mombers C., de Gier J., Demel R. A., van Deenen L. L. Spectrin-phospholipid interaction. A monolayer study. Biochim Biophys Acta. 1980 Dec 2;603(1):52–62. doi: 10.1016/0005-2736(80)90390-9. [DOI] [PubMed] [Google Scholar]
  19. Op den Kamp J. A. Lipid asymmetry in membranes. Annu Rev Biochem. 1979;48:47–71. doi: 10.1146/annurev.bi.48.070179.000403. [DOI] [PubMed] [Google Scholar]
  20. Pagano R. E., Martin O. C., Schroit A. J., Struck D. K. Formation of asymmetric phospholipid membranes via spontaneous transfer of fluorescent lipid analogues between vesicle populations. Biochemistry. 1981 Aug 18;20(17):4920–4927. doi: 10.1021/bi00520a018. [DOI] [PubMed] [Google Scholar]
  21. Palek J., Liu S. C., Snyder L. M. Metabolic dependence of protein arrangement in human erythrocyte membranes. I. Analysis of spectrin-rich complexes in ATP-depleted red cells. Blood. 1978 Mar;51(3):385–395. [PubMed] [Google Scholar]
  22. Patel V. P., Fairbanks G. Spectrin phosphorylation and shape change of human erythrocyte ghosts. J Cell Biol. 1981 Feb;88(2):430–440. doi: 10.1083/jcb.88.2.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Quist E. E., Reece K. L. The role of diphosphatidylinositol in erythrocyte membrane shape regulation. Biochem Biophys Res Commun. 1980 Aug 14;95(3):1023–1030. doi: 10.1016/0006-291x(80)91575-2. [DOI] [PubMed] [Google Scholar]
  24. Rousselet A., Guthmann C., Matricon J., Bienvenue A., Devaux P. F. Study of the transverse diffusion of spin labeled phospholipids in biological membranes. I. Human red bloods cells. Biochim Biophys Acta. 1976 Mar 19;426(3):357–371. doi: 10.1016/0005-2736(76)90382-5. [DOI] [PubMed] [Google Scholar]
  25. Schwoch G., Passow H. Preparation and properties of human erythrocyte ghosts. Mol Cell Biochem. 1973 Dec 15;2(2):197–218. doi: 10.1007/BF01795474. [DOI] [PubMed] [Google Scholar]
  26. Sheetz M. P., Singer S. J. Biological membranes as bilayer couples. A molecular mechanism of drug-erythrocyte interactions. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4457–4461. doi: 10.1073/pnas.71.11.4457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sheetz M. P., Singer S. J. On the mechanism of ATP-induced shape changes in human erythrocyte membranes. I. The role of the spectrin complex. J Cell Biol. 1977 Jun;73(3):638–646. doi: 10.1083/jcb.73.3.638. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tanaka K. I., Ohnishi S. Heterogeneity in the fluidity of intact erythrocyte membrane and its homogenization upon hemolysis. Biochim Biophys Acta. 1976 Mar 5;426(2):218–231. doi: 10.1016/0005-2736(76)90333-3. [DOI] [PubMed] [Google Scholar]
  29. Verkleij A. J., Zwaal R. F., Roelofsen B., Comfurius P., Kastelijn D., van Deenen L. L. The asymmetric distribution of phospholipids in the human red cell membrane. A combined study using phospholipases and freeze-etch electron microscopy. Biochim Biophys Acta. 1973 Oct 11;323(2):178–193. doi: 10.1016/0005-2736(73)90143-0. [DOI] [PubMed] [Google Scholar]
  30. Zwaal R. F., Roelofsen B., Comfurius P., van Deenen L. L. Organization of phospholipids in human red cell membranes as detected by the action of various purified phospholipases. Biochim Biophys Acta. 1975 Sep 16;406(1):83–96. doi: 10.1016/0005-2736(75)90044-9. [DOI] [PubMed] [Google Scholar]
  31. van Meer G., Op den Kamp J. A. Transbilayer movement of various phosphatidylcholine species in intact human erythrocytes. J Cell Biochem. 1982;19(2):193–204. doi: 10.1002/jcb.240190209. [DOI] [PubMed] [Google Scholar]

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