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Patch-Clamped Liposomes

Recording Reconstituted Ion Channels
  • David W. Tank
  • Christopher Miller

Abstract

Ion-channel reconstitution and patch recording offer different but complementary experimental methods. Both can be used to help provide a molecular understanding of how ion transport activity is controlled by protein structure and regulation. On the one hand, through use of reconstitution techniques, an ion channel can be isolated, purified, and/or chemically modified and reinserted into a lipid bilayer. The result is a well-characterized membrane system in which biochemical properties likely to be important in regulating transport—subunit stoichiometry, phosphorylation, methylation, lipid composition, etc.—can be monitored and controlled. On the other hand, patch recording techniques can provide the necessary sensitivity in ion transport measurements for discrimination of changes in single channel activity that are effected by structural and regulatory alterations, and the measurements can be done in an experimental geometry that allows controlled access to the membrane of bath-soluble chemical messengers and proteins.

Keywords

Planar Lipid Bilayer Small Unilamellar Vesicle Patch Recording Single Bilayer Nicotinic AChR 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Boheim, G., Hanke, W., Barrantes, F. J., Eibl, H., Sakmann, B., Fels, G., and Maelicke, A., 1981, Agonist-activated ionic channels in acetylcholine receptor reconstituted into planar lipid bilayers, Proc. Natl. Acad. Sci. U.S.A. 78:3586–3590.PubMedCrossRefGoogle Scholar
  2. Changeux, J. P., Heidmann, T., Popot, J. L., and Sobel, A., 1979, Reconstitution of a functional acetylcholine regulator under defined conditions, FEBS Lett. 105:181–187.PubMedCrossRefGoogle Scholar
  3. Cuppoletti, J., Mayhew, E., Zobel, C. R., and Jung, C. Y., 1981, Erythrosomes: Large proteoliposomes derived from crosslinked human erythrocyte cytoskeletons and exogenous lipid, Proc. Natl. Acad. Sci. U.S.A. 78:2786–2790.PubMedCrossRefGoogle Scholar
  4. Hamill, O. P., Marty, A., Neher, E., Sakmann, B., and Sigworth, F. J., 1981, Improved patch-clamp techniques for high-resolution current recordings from cells and cell-free membrane patches, Pflügers Arch. 391:85–100.PubMedCrossRefGoogle Scholar
  5. Herrmann, S. H., and Mescher, M. F., 1981, Secondary cytolytic T lymphocyte stimulation by purified H-2K in liposomes, Proc. Natl. Acad. Sci. U.S.A. 78:2488–2492.PubMedCrossRefGoogle Scholar
  6. Horn, R., and Patlak, J., 1980, Single channel currents from excised patches of muscle membrane, Proc. Natl. Acad. Sci. U.S.A. 77:6930–6934.PubMedCrossRefGoogle Scholar
  7. Hub, H. H., Zimmermann, U., and Ringsdorf, H., 1982, Preparation of large unilamellar vesicles, FEBS Lett. 140:254–256.CrossRefGoogle Scholar
  8. Huganir, R. L., and Racker, E., 1982, Properties of proteoliposomes reconstituted with acetylcholine receptor from Torpedo californica, J. Biol. Chem. 257:9372–9378.Google Scholar
  9. Huganir, R. L., Schell, M. A., and Racker, E., 1979, Reconstitution of the purified acetylcholine receptor from Torpedo californica, FEBS Lett. 108:155–160.CrossRefGoogle Scholar
  10. Kagawa, Y., and Racker, E., 1971, Partial resolution of enzymes catalyzing oxidative phosphorylation. 25.Google Scholar
  11. Reconstitution of vesicles catalyzing 32Pi-adenosine-triphosphate exchange, J. Biol. Chem. 246:5477–5487.Google Scholar
  12. Kasahara, M., and Hinkle, P. C., 1977, Reconstitution and purification of the D-glucose transporter from human erythrocytes, J. Biol. Chem. 252:7384–7390.PubMedGoogle Scholar
  13. Krishtal, O. A., and Pidoplichko, V. I., 1980, A receptor for protons in the nerve cell membrane, Neuroscience 5:2325–2327.PubMedCrossRefGoogle Scholar
  14. Lindstrom, J., Anholt, R., Einarson, B., Engel, A., Osame, M., and Montai, M., 1980, Purification of acetylcholine receptors, reconstitution into lipid bilayers, and study of agonist-induced cation channel regulation,J. Biol. Chem. 255:8340–8350.PubMedGoogle Scholar
  15. Miller, C., 1982, Open-state substructure of single chloride channels from Torpedo electroplax, Phil. Trans. Roy. Soc. Lond. B 299:401–411.CrossRefGoogle Scholar
  16. Miller, C., 1982, Reconstitution of ion channels in planar bilayer membranes: A five year progress report, Comm. Mol. Cell Biophys. 1:413–428.Google Scholar
  17. Miller, C., and Racker, E., 1979, Reconstitution of membrane transport functions, in: The Receptors: A Comprehensive Treatise, Vol. 1 (R. D. O’Brien, ed.), pp. 1–31, Plenum Press, New York.Google Scholar
  18. Miller, C., and White, M. M., 1980, A voltage-dependent chloride conductance from Torpedo electroplax membrane, Ann. N.Y. Acad. Sci. 341:534–551.PubMedCrossRefGoogle Scholar
  19. Nelson, N., Anholt, R., Lindstrom, J., and Montai, M., 1980, Reconstitution of purified acetylcholine receptors with functional ion channels in planar lipid bilayers, Proc. Natl. Acad. Sci. U.S.A. 77:3057–3061.PubMedCrossRefGoogle Scholar
  20. Neubig, R. R., Boyd, N. D., and Cohen, J. B., 1982, Conformations of Torpedo acetylcholine receptor associated with ion transport and desensitization, Biochemistry 21:3460–3467.PubMedCrossRefGoogle Scholar
  21. Papahadjopoulos, D., Vail, W. J., Jacobson, K., and Poste, G., 1975, Cochleate lipid cylinders: Formation by fusion of unilamellar lipid vesicles, Biochim. Biophys. Acta 394:483–491.PubMedCrossRefGoogle Scholar
  22. Pick, U., 1981, Liposomes with large trapping capacity prepared by freezing and thawing of sonicated phospholipid mixtures, Arch. Biochem. Biophys. 212:186–194.PubMedCrossRefGoogle Scholar
  23. Reeves, J. P., and Dowben, R. M., 1969, Formation and properties of thin-walled phospholipid vesicles,J. Cell. Physiol. 73:49–60.PubMedCrossRefGoogle Scholar
  24. Sakmann, B., Patlak, J., and Neher, E., 1980, Single acetylcholine-activated channels show burst-kinetics in presence of desensitizing concentrations of agonist, Nature 286:71–73.PubMedCrossRefGoogle Scholar
  25. Schindler, H., and Quast, U., 1980, Functional acetylcholine receptor from Torpedo marmorata in planar membranes, Proc. Natl. Acad. Sci. U.S.A. 77:3052–3056.PubMedCrossRefGoogle Scholar
  26. Szoka, F., and Papahadjopoulos, D., 1980, Comparative properties and methods of preparation of lipid vesicles (liposomes), Annu. Rev. Biophys. Bioeng. 9:467–508.PubMedCrossRefGoogle Scholar
  27. Talvenheimo, J. A., Tamkun, M. M., and Catterall, W. A., 1982, Reconstitution of a functional mammalian sodium channel from partially purified components, Soc. Neurosci. Abstr. 8:727.Google Scholar
  28. Tank, D. W., Miller, C., and Webb, W. W., 1982, Isolated-patch recording from liposomes containing functionally reconstituted chloride channels from Torpedo electroplax, Proc. Natl. Acad. Sci. U.S.A. 79:7749–7753.PubMedCrossRefGoogle Scholar
  29. Vail, W. J., and Stollery, J. G., 1979, Phase changes of cardiolipin vesicles mediated by different cations, Biochim. Biophys. Acta 551:74–84.PubMedCrossRefGoogle Scholar
  30. Walker, J. W., McNamee, M. G., Pasquale, E., Cash, D. J., and Hess, G. P., 1981, Acetylcholine receptor inactivation in Torpedo californica electroplax membrane vesicles. Detection of two processes in the millisecond and second time regions, Biochem. Biophys. Res. Commun. 100:86–90.PubMedCrossRefGoogle Scholar
  31. Weigele, J. B., and Barchi, R. L., 1982, Functional reconstitution of the purified sodium channel protein from rat sarcolemma, Proc. Natl. Acad. Sci. U.S.A. 79:3651–3655.PubMedCrossRefGoogle Scholar
  32. Wu, W. C. S., and Raftery, M. A., 1979, Carbamylcholine-induced rapid cation efflux from reconstituted vesicles containing purified acetylcholine receptor, Biochem. Biophys. Res. Commun. 89:26–35.PubMedCrossRefGoogle Scholar
  33. Yellen, G., 1982, Single Ca++-activated nonselective cation channels in neuroblastoma, Nature 296:357–359.PubMedCrossRefGoogle Scholar
  34. Zimmerman, U., and Scheurich, P., 1981, Fusion of Avena sativa mesophyll cell protoplasts by electrical breakdown, Biochim. Biophys. Acta 641:160–165.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1983

Authors and Affiliations

  • David W. Tank
    • 1
  • Christopher Miller
    • 2
  1. 1.Department of PhysicsCornell UniversityIthacaUSA
  2. 2.Graduate Department of BiochemistryBrandeis UniversityWalthamUSA

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