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. 1977 Aug 1;74(2):531–546. doi: 10.1083/jcb.74.2.531

Adhesion of phospholipid vesicles to Chinese hamster fibroblasts: Role of cell surface proteins

RE Pagano, M Takeichi
PMCID: PMC2110067  PMID: 407233

Abstract

The adhesion of artificially generated lipid membrane vesicles to Chinese hamster V79 fibroblasts in suspension was used as a model system for studying membrane interactions. Below their gel-liquid crystalline phase transition temperature, vesicles comprised of dipalmitoyl lecithin (DPL) or dimyristoyl lecithin (DML) absorbed to the surfaces of EDTA- dissociated cells. These adherent vesicles could not be removed by repeated washings of the treated cells but could be released into the medium by treatment with trypsin. EM autoradiographic studies of cells treated with[(3)H]DML or [(3)H]DPL vesicles showed that most of the radioactive lipids were confined to the cell periphery. Scanning electron microscopy and fluorescence microscopy further confirmed the presence of adherent vesicles at the cell surface. Adhesion of DML or DPL vesicles to EDTA-dissociated cells modified the lactoperoxidase-catalyzed iodination pattern of the cell surface proteins; the inhibition of labeling of two proteins with an approximately 60,000- dalton mol wt was particularly evident. Incubation of cells wit h (3)H-lipid vesicles followed by sodium dodecyl sulfate (SDS)- polyacrylamide gel electrophoresis showed that some of the (3)H-lipid migrated preferentially with these approximately 60,000-mol wt proteins. Studies of the temperature dependence of vesicle uptake and subsequent release by trypsin showed that DML or DPL vesicle adhesion to EDTA- dissociated cells increased with decreasing temperatures. In contrast, cells trypsinized before incubation with vesicles showed practically no temperature dependence of vesicle uptake. These results suggest two pathways for adhesion of lipid vesicles to the cell surface-a temperature-sensitive one involving cell surface proteins, and a temperature-independent one. These findings are discussed in terms of current models for cell-cell interactions.

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Selected References

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  1. Alderson J. C., Green C. Enrichment of lymphocytes with cholesterol and its effect on lymphocyte activation. FEBS Lett. 1975 Apr 1;52(2):208–211. doi: 10.1016/0014-5793(75)80807-6. [DOI] [PubMed] [Google Scholar]
  2. Awasthi Y. C., Chuang T. F., Keenan T. W., Crane F. L. Tightly bound cardiolipin in cytochrome oxidase. Biochim Biophys Acta. 1971 Jan 12;226(1):42–52. doi: 10.1016/0005-2728(71)90176-9. [DOI] [PubMed] [Google Scholar]
  3. Bangham A. D. Lipid bilayers and biomembranes. Annu Rev Biochem. 1972;41:753–776. doi: 10.1146/annurev.bi.41.070172.003541. [DOI] [PubMed] [Google Scholar]
  4. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  5. Bruckdorfer K. R., Demel R. A., De Gier J., van Deenen L. L. The effect of partial replacements of membrane cholesterol by other steroids on the osmotic fragility and glycerol permeability of erythrocytes. Biochim Biophys Acta. 1969 Jul 15;183(2):334–345. doi: 10.1016/0005-2736(69)90089-3. [DOI] [PubMed] [Google Scholar]
  6. CARO L. G., VAN TUBERGEN R. P., KOLB J. A. High-resolution autoradiography. I. Methods. J Cell Biol. 1962 Nov;15:173–188. doi: 10.1083/jcb.15.2.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chen J. S., Del Fa A., Di Luzio A., Calissano P. Liposome-induced morphological differentiation of murine neuroblastoma. Nature. 1976 Oct 14;263(5578):604–606. doi: 10.1038/263604a0. [DOI] [PubMed] [Google Scholar]
  8. Curtis A. S., Chandler C., Picton N. Cell surface lipids and adhesion. III. The effects on cell adhesion of changes in plasmalemmal lipids. J Cell Sci. 1975 Aug;18(3):375–384. doi: 10.1242/jcs.18.3.375. [DOI] [PubMed] [Google Scholar]
  9. Grant C. W., McConnell H. M. Fusion of phospholipid vesicles with viable Acholeplasma laidlawii. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1238–1240. doi: 10.1073/pnas.70.4.1238. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hauser H. O. The effect of ultrasonic irradiation on the chemical structure of egg lecithin. Biochem Biophys Res Commun. 1971 Nov;45(4):1049–1055. doi: 10.1016/0006-291x(71)90443-8. [DOI] [PubMed] [Google Scholar]
  11. Helenius A., Simons K. Solubilization of membranes by detergents. Biochim Biophys Acta. 1975 Mar 25;415(1):29–79. doi: 10.1016/0304-4157(75)90016-7. [DOI] [PubMed] [Google Scholar]
  12. Huang C., Charlton J. P. Studies on the state of phosphatidylcholine molecules before and after ultrasonic and gel-filtration treatments. Biochem Biophys Res Commun. 1972 Feb 25;46(4):1660–1666. doi: 10.1016/0006-291x(72)90800-5. [DOI] [PubMed] [Google Scholar]
  13. Huang C. Studies on phosphatidylcholine vesicles. Formation and physical characteristics. Biochemistry. 1969 Jan;8(1):344–352. doi: 10.1021/bi00829a048. [DOI] [PubMed] [Google Scholar]
  14. Huang L., Pagano R. E. Interaction of phospholipid vesicles with cultured mammalial cells. I. Characteristics of uptake. J Cell Biol. 1975 Oct;67(1):38–48. doi: 10.1083/jcb.67.1.38. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hubbard A. L., Cohn Z. A. The enzymatic iodination of the red cell membrane. J Cell Biol. 1972 Nov;55(2):390–405. doi: 10.1083/jcb.55.2.390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Inbar M., Shinitzky M. Increase of cholesterol level in the surface membrane of lymphoma cells and its inhibitory effect on ascites tumor development. Proc Natl Acad Sci U S A. 1974 May;71(5):2128–2130. doi: 10.1073/pnas.71.5.2128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Magee W. E., Goff C. W., Schoknecht J., Smith M. D., Cherian K. The interaction of cationic liposomes containing entrapped horseradish peroxidase with cells in culture. J Cell Biol. 1974 Nov;63(2 Pt 1):492–504. doi: 10.1083/jcb.63.2.492. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Martin F. J., MacDonald R. C. Lipid vesicle-cell interactions. III. Introduction of a new antigenic determinant into erythrocyte membranes. J Cell Biol. 1976 Sep;70(3):515–526. doi: 10.1083/jcb.70.3.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. PUCK T. T., CIECIURA S. J., ROBINSON A. Genetics of somatic mammalian cells. III. Long-term cultivation of euploid cells from human and animal subjects. J Exp Med. 1958 Dec 1;108(6):945–956. doi: 10.1084/jem.108.6.945. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Pagano R. E., Huang L. Interaction of phospholipid vesicles with cultured mammalian cells. II. Studies of mechanism. J Cell Biol. 1975 Oct;67(1):49–60. doi: 10.1083/jcb.67.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pagano R. E., Huang L., Wey C. Interaction of phospholipid vesicles with cultured mammalian cells. Nature. 1974 Nov 8;252(5479):166–167. doi: 10.1038/252166a0. [DOI] [PubMed] [Google Scholar]
  22. Pagano R. E., Ozato K., Ruysschaert J. M. Intracellular distribution of lipophilic fluorescent probes in mammalian cells. Biochim Biophys Acta. 1977 Mar 17;465(3):661–666. doi: 10.1016/0005-2736(77)90282-6. [DOI] [PubMed] [Google Scholar]
  23. Papahadjopoulos D., Poste G., Schaeffer B. E. Fusion of mammalian cells by unilamellar lipid vesicles: inflluence of lipid surface charge, fluidity and cholesterol. Biochim Biophys Acta. 1973 Sep 27;323(1):23–42. doi: 10.1016/0005-2736(73)90429-x. [DOI] [PubMed] [Google Scholar]
  24. Phillips D. R., Morrison M. Exposed protein on the intact human erythrocyte. Biochemistry. 1971 May 11;10(10):1766–1771. doi: 10.1021/bi00786a006. [DOI] [PubMed] [Google Scholar]
  25. Poste G., Papahadjopoulos D. Lipid vesicles as carriers for introducing materials into cultured cells: influence of vesicle lipid composition on mechanism(s) of vesicle incorporation into cells. Proc Natl Acad Sci U S A. 1976 May;73(5):1603–1607. doi: 10.1073/pnas.73.5.1603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Rahman Y. E., Wright B. J. Liposomes containing chelating agents. Cellular penetration and a possible mechanism of metal removal. J Cell Biol. 1975 Apr;65(1):112–122. doi: 10.1083/jcb.65.1.112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stambrook P. J., Sisken J. E. Induced changes in the rates of uridine- 3 H uptake and incorporation during the G 1 and S periods of synchronized Chinese hamster cells. J Cell Biol. 1972 Mar;52(3):514–525. doi: 10.1083/jcb.52.3.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Weinstein J. N., Yoshikami S., Henkart P., Blumenthal R., Hagins W. A. Liposome-cell interaction: transfer and intracellular release of a trapped fluorescent marker. Science. 1977 Feb 4;195(4277):489–492. doi: 10.1126/science.835007. [DOI] [PubMed] [Google Scholar]
  29. Weisman R. A., Korn E. D. Uptake of fatty acids by Acanthamoeba. Biochim Biophys Acta. 1966 Apr 4;116(2):229–242. doi: 10.1016/0005-2760(66)90006-3. [DOI] [PubMed] [Google Scholar]
  30. Weissmann G., Bloomgarden D., Kaplan R., Cohen C., Hoffstein S., Collins T., Gotlieb A., Nagle D. A general method for the introduction of enzymes, by means of immunoglobulin-coated liposomes, into lysosomes of deficient cells. Proc Natl Acad Sci U S A. 1975 Jan;72(1):88–92. doi: 10.1073/pnas.72.1.88. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wetzel M. G., Korn E. D. Phagocytosis of latex beads by Acahamoeba castellanii (Neff). 3. Isolation of the phagocytic vesicles and their membranes. J Cell Biol. 1969 Oct;43(1):90–104. doi: 10.1083/jcb.43.1.90. [DOI] [PMC free article] [PubMed] [Google Scholar]

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