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. 1981 May;78(5):2786–2790. doi: 10.1073/pnas.78.5.2786

Erythrosomes: large proteoliposomes derived from crosslinked human erythrocyte cytoskeletons and exogenous lipid.

J Cuppoletti, E Mayhew, C R Zobel, C Y Jung
PMCID: PMC319442  PMID: 6789323

Abstract

Large (3-micrometers diameter) mechanically stable proteoliposomes (erythrosomes) were prepared in good yield by coating crosslinked erythrocyte cytoskeletons with phosphatidylcholine. The erythrosomes consist of the polypeptides designated band 1, 2, 3, 4.1 + 4.2, and 5 (less than 4% of the endogenous lipid) and enough added lipid to form a bilayer coating the surface. Electron microscopy shows only the large proteoliposomes in sealed preparations. The trapping of bovine serum albumin, mannitol, sucrose, glucose, cytosine arabinoside, and sodium in the erythrosomes was demonstrated, yielding an apparent volume of up to 100 liters/mol of phospholipid. This preparation possesses an effective diffusion barrier to glucose, sucrose, and sodium ion with half-equilibration times of 34, 29, and 170 hr, respectively. The results of the present study suggest that erythrosomes may be useful for membrane transport protein reconstitution and encapsulation systems.

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Selected References

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  1. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Cecchini G., Payne G. S., Oxender D. L. Reconstitution of neutral amino acid transport from partially purified membrane components from Ehrlich ascites tumor cells. J Supramol Struct. 1977;7(3-4):481–487. doi: 10.1002/jss.400070317. [DOI] [PubMed] [Google Scholar]
  4. DODGE J. T., MITCHELL C., HANAHAN D. J. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963 Jan;100:119–130. doi: 10.1016/0003-9861(63)90042-0. [DOI] [PubMed] [Google Scholar]
  5. Enoch H. G., Strittmatter P. Formation and properties of 1000-A-diameter, single-bilayer phospholipid vesicles. Proc Natl Acad Sci U S A. 1979 Jan;76(1):145–149. doi: 10.1073/pnas.76.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fahey P. F., Webb W. W. Lateral diffusion in phospholipid bilayer membranes and multilamellar liquid crystals. Biochemistry. 1978 Jul 25;17(15):3046–3053. doi: 10.1021/bi00608a016. [DOI] [PubMed] [Google Scholar]
  7. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  8. Ji T. H., Kiehm D. J., Middaugh C. R. Presence of spectrin tetramer on the erythrocyte membrane. J Biol Chem. 1980 Apr 10;255(7):2990–2993. [PubMed] [Google Scholar]
  9. Kasahara M., Hinkle P. C. Reconstitution and purification of the D-glucose transporter from human erythrocytes. J Biol Chem. 1977 Oct 25;252(20):7384–7390. [PubMed] [Google Scholar]
  10. Marinetti G. V., Baumgarten R., Sheeley D., Gordesky S. Cross-linking of phospholipids to proteins in the erythrocyte membrane. Biochem Biophys Res Commun. 1973 Jul 2;53(1):302–308. doi: 10.1016/0006-291x(73)91434-4. [DOI] [PubMed] [Google Scholar]
  11. Mombers C., van Dijck P. W., van Deenen L. L., de Gier J., Verkleij A. J. The interaction of spectrin - actin and synthetic phospholipids. Biochim Biophys Acta. 1977 Oct 17;470(2):152–160. doi: 10.1016/0005-2736(77)90096-7. [DOI] [PubMed] [Google Scholar]
  12. Montelaro R. C., Rueckert R. R. Radiolabeling of proteins and viruses in vitro by acetylation with radioactive acetic anhydride. J Biol Chem. 1975 Feb 25;250(4):1413–1421. [PubMed] [Google Scholar]
  13. Racker E. Reconstitution of a calcium pump with phospholipids and a purified Ca ++ - adenosine triphosphatase from sacroplasmic reticulum. J Biol Chem. 1972 Dec 25;247(24):8198–8200. [PubMed] [Google Scholar]
  14. Shapiro A. L., Viñuela E., Maizel J. V., Jr Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun. 1967 Sep 7;28(5):815–820. doi: 10.1016/0006-291x(67)90391-9. [DOI] [PubMed] [Google Scholar]
  15. Sheetz M. P. Integral membrane protein interaction with Triton cytoskeletons of erythrocytes. Biochim Biophys Acta. 1979 Oct 19;557(1):122–134. doi: 10.1016/0005-2736(79)90095-6. [DOI] [PubMed] [Google Scholar]
  16. Sheetz M. P., Sawyer D. Triton shells of intact erythrocytes. J Supramol Struct. 1978;8(4):399–412. doi: 10.1002/jss.400080403. [DOI] [PubMed] [Google Scholar]
  17. Szoka F., Jr, Papahadjopoulos D. Procedure for preparation of liposomes with large internal aqueous space and high capture by reverse-phase evaporation. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4194–4198. doi: 10.1073/pnas.75.9.4194. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Thompson N. L., Axelrod D. Reduced lateral mobility of a fluorescent lipid probe in cholesterol-depleted erythrocyte membrane. Biochim Biophys Acta. 1980 Mar 27;597(1):155–165. doi: 10.1016/0005-2736(80)90159-5. [DOI] [PubMed] [Google Scholar]
  19. Volsky D. J., Cabantchik Z. I., Beigel M., Loyter A. Implantation of the isolated human erythrocyte anion channel into plasma membranes of Friend erythroleukemic cells by use of Sendai virus envelopes. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5440–5444. doi: 10.1073/pnas.76.11.5440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Yu J., Fischman D. A., Steck T. L. Selective solubilization of proteins and phospholipids from red blood cell membranes by nonionic detergents. J Supramol Struct. 1973;1(3):233–248. doi: 10.1002/jss.400010308. [DOI] [PubMed] [Google Scholar]

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