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Measurement of Ca2+ Flux Through Ins(1,4,5)P3 Receptor-Ca2+ Channels in Lipid Bilayers (“Dip-Tip” and “Schindler” Methodology)

  • Edwin C. Thrower
Part of the Methods in Molecular Biology™ book series (MIMB, volume 114)

Abstract

The inositol (1,4,5)trisphosphate [Ins(1,4,5)P3] receptor [Ins(1,4,5)P3-sensitive-Ca2+ channel] is an intracellular Ca2+ release channel located within the membrane of the smooth endoplasmic reticulum (ER). The study of kinetics and regulation of the Ins(1,4,5)P3 receptor is greatly complicated by problems of the heterogeneity of its environment (such as vesicle size or the number of receptors per unit area of membrane), and difficulties arise in achieving tight control of the ionic composition on the luminal side of the channel. These problems can be overcome, to a certain extent, by monitoring single-channel characteristics of the Ins(1,4,5)P3 receptor (such as channel unit conductance and open and closed times) in artificial planar lipid bilayers and relating this information to that obtained from various other studies.

Keywords

Critical Micellar Concentration Soybean Lecithin Silicone Grease Teflon Film Vesicle Suspension 
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.

References

  1. 1.
    Ehrlich, B. E. (1992) Planar lipid bilayers on patch pipettes: Bilayer formation and ion channel incorporation. Methods Enzymol. 207, 463–471.PubMedCrossRefGoogle Scholar
  2. 2.
    Schindler, H. and Feher, G. (1976) Branched molecular lipid membranes. Biophys. J. 16, 109–111.CrossRefGoogle Scholar
  3. 3.
    Schindler, H. (1980) Formation of planar lipid bilayers from artificial or native membrane vesicles. FEBS Lett. 122, 77–79.PubMedCrossRefGoogle Scholar
  4. 4.
    Hamill, O. P., Marty, A., Neher, E., Sakmann, B., and Sigworth, F. J. (1981) Improved patch-clamp technique for high resolution current recording from cells and cell-free membrane patches. Pflügers Arch. 391, 85–100.PubMedCrossRefGoogle Scholar
  5. 5.
    Sakmann, B. and Neher, E. (1983) Single Channel Recording. Plenum, New York.Google Scholar
  6. 6.
    Coronado, R. and Latorre, R. (1983) Formation of phospholipid bilayers on patch-clamp pipettes. Biophys. J. 43, 231–236.PubMedCrossRefGoogle Scholar
  7. 7.
    Schuerholz, T. and Schindler, H. (1983) Formation of lipid-protein bilayers by micropipette-guided contact of two monolayers. FEBS Lett. 152, 187–190.CrossRefGoogle Scholar
  8. 8.
    Suarez-Izla, B. A., Wan, K., Lindstrom, J., and Montal, M. (1983) Single channel recordings from purified acetylcholine receptors reconstituted in bilayers formed at the tip of patch pipettes. Biochemistry 22, 2319–2323.CrossRefGoogle Scholar
  9. 9.
    Wilmsen, U., Methfessel, C., Hanke, W., and Boheim, G. (1983) Channel current fluctuation studies with solvent free planar lipid bilayers using Neher-Sakmann pipettes, in Physical Chemistry of Transmembrane Ion Motion (Spach, G., ed.), Elsevier, Amsterdam, pp. 479–485.Google Scholar
  10. 10.
    Hanke, W., Methfessel, C., Wilmsen, U., and Boheim, G. (1984) Ion channel reconstitution into planar lipid bilayers on glass pipettes. Biochem. Bioeng. J. 12, 329–339.CrossRefGoogle Scholar
  11. 11.
    Langmuir, I. and Waugh, D. F. (1938) The adsorption of proteins at oil-water interfaces and artificial protein-lipid membranes. J. Gen. Physiol. 21, 745–755.PubMedCrossRefGoogle Scholar
  12. 12.
    Takagi, M., Azuma, K., and Kishimoto, U. (1965) A new method for the formation of bilayer membranes in aqueous solutions. Annu. Rep. Biol. Fac. Sci. Osaka 13, 107–110.Google Scholar
  13. 13.
    Montal, M. and Mueller, P. (1972) Formation of bimolecular membranes from lipid monolayers and a study of their electrical properties. Proc. Natl. Acad. Sci. USA 69, 3561–3566.PubMedCrossRefGoogle Scholar
  14. 14.
    Danoff, S. K., Supattapone, S., and Snyder, S. H. (1988) Characterisation of a membrane protein from brain mediating the inhibition of inositol 1,4,5-trisphosphate receptor-binding by calcium. Biochem. J. 254, 701–705.PubMedGoogle Scholar
  15. 15.
    Bezprozvanny, I. and Ehrlich, B. E. (1993) ATP modulates the function of inosi-tol 1,4,5 trisphosphate-gated channels at two sites. Neuron 10, 1175–1184.PubMedCrossRefGoogle Scholar
  16. 16.
    Bezprozvanny, I., Watras, J., and Ehrlich, B. E. (1991) Bell-shaped calcium response curves for the Ins(1,4,5)P3-and calcium-gated channels from endoplasmic reticulum. Nature 351, 751–754.PubMedCrossRefGoogle Scholar
  17. 17.
    Bezprozvanny, I. and Ehrlich, B. E. (1994) Inositol 1,4,5-trisphosphate (InsP3)-gated Ca channels from cerebellum: Conduction properties for divalent cations and regulation by intraluminal calcium. J. Gen. Physiol. 104, 821–856.PubMedCrossRefGoogle Scholar
  18. 18.
    Ferris, C. D., Huganir, R. L., Supattapone, S., and Snyder, S. H. (1989) Purified inositol 1,4,5-trisphosphate receptor mediates calcium flux in reconstituted lipid vesicles. Nature (Lond.) 342, 87–89.CrossRefGoogle Scholar
  19. 19.
    Nakade, S., Rhee, S. K., Hamanaka, H., and Miksohiba, K. (1994) Cyclic AMP-dependent phosphorylation of an immunoaffinity-purified homotetrameric inositol 1,4,5-trisphosphate receptor (Type I) increases Ca2+ flux in reconstituted lipid vesicles. J. Biol. Chem. 269, 6735–6742.PubMedGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1999

Authors and Affiliations

  • Edwin C. Thrower
    • 1
  1. 1.University of East AngliaNorwichUK

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