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. 1991 Aug;60(2):380–388. doi: 10.1016/S0006-3495(91)82063-X

Voltage-dependent conductance for alamethicin in phospholipid vesicles. A test for the mechanism of gating.

S J Archer 1, D S Cafiso 1
PMCID: PMC1260074  PMID: 1717015

Abstract

The ion currents induced by alamethicin were investigated in unilamellar vesicles using electron paramagnetic resonance probe techniques. The peptide induced currents were examined as a function of the membrane bound peptide concentration, and as a function of the transmembrane electrical potential. Because of the favorable partitioning of alamethicin to membranes and the large membrane area to aqueous volume in vesicle suspensions, these measurements could be carried out under conditions where all the alamethicin was membrane bound. Over the concentration range examined, the peptide induced conductances increased approximately with the fourth power of the membrane bound peptide concentration, indicating a channel molecularity of four. When the alamethicin induced currents were examined as a function of voltage, they exhibited a superlinear behavior similar to that seen in planar bilayers. Evidence for the voltage-dependent conduction of alamethicin was also observed in the time dependence of vesicle depolarization. These observations indicate that the voltage-dependent behavior of alamethicin can occur in the absence of a voltage-dependent phase partitioning. That is, a voltage-dependent conformational rearrangement for membrane bound alamethicin leads to a voltage-dependent activity.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Archer S. J., Ellena J. F., Cafiso D. S. Dynamics and aggregation of the peptide ion channel alamethicin. Measurements using spin-labeled peptides. Biophys J. 1991 Aug;60(2):389–398. doi: 10.1016/S0006-3495(91)82064-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  3. Bruggemann E. P., Kayalar C. Determination of the molecularity of the colicin E1 channel by stopped-flow ion flux kinetics. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4273–4276. doi: 10.1073/pnas.83.12.4273. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cafiso D. S. Electron paramagnetic resonance methods for measuring pH gradients, transmembrane potentials, and membrane dynamics. Methods Enzymol. 1989;172:331–345. doi: 10.1016/s0076-6879(89)72022-x. [DOI] [PubMed] [Google Scholar]
  5. Cafiso D. S., Hubbell W. L. EPR determination of membrane potentials. Annu Rev Biophys Bioeng. 1981;10:217–244. doi: 10.1146/annurev.bb.10.060181.001245. [DOI] [PubMed] [Google Scholar]
  6. Cafiso D. S., Hubbell W. L. Estimation of transmembrane pH gradients from phase equilibria of spin-labeled amines. Biochemistry. 1978 Sep 5;17(18):3871–3877. doi: 10.1021/bi00611a030. [DOI] [PubMed] [Google Scholar]
  7. Cascio M., Wallace B. A. Conformation of alamethicin in phospholipid vesicles: implications for insertion models. Proteins. 1988;4(2):89–98. doi: 10.1002/prot.340040203. [DOI] [PubMed] [Google Scholar]
  8. Eisenberg M., Hall J. E., Mead C. A. The nature of the voltage-dependent conductance induced by alamethicin in black lipid membranes. J Membr Biol. 1973 Dec 31;14(2):143–176. doi: 10.1007/BF01868075. [DOI] [PubMed] [Google Scholar]
  9. Flewelling R. F., Hubbell W. L. Hydrophobic ion interactions with membranes. Thermodynamic analysis of tetraphenylphosphonium binding to vesicles. Biophys J. 1986 Feb;49(2):531–540. doi: 10.1016/S0006-3495(86)83663-3. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hall J. E., Vodyanoy I., Balasubramanian T. M., Marshall G. R. Alamethicin. A rich model for channel behavior. Biophys J. 1984 Jan;45(1):233–247. doi: 10.1016/S0006-3495(84)84151-X. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lau A. L., Chan S. I. Voltage-induced formation of alamethicin pores in lecithin bilayer vesicles. Biochemistry. 1976 Jun 15;15(12):2551–2555. doi: 10.1021/bi00657a010. [DOI] [PubMed] [Google Scholar]
  12. Montal M., Mueller P. Formation of bimolecular membranes from lipid monolayers and a study of their electrical properties. Proc Natl Acad Sci U S A. 1972 Dec;69(12):3561–3566. doi: 10.1073/pnas.69.12.3561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Rizzo V., Stankowski S., Schwarz G. Alamethicin incorporation in lipid bilayers: a thermodynamic study. Biochemistry. 1987 May 19;26(10):2751–2759. doi: 10.1021/bi00384a015. [DOI] [PubMed] [Google Scholar]
  14. SINGLETON W. S., GRAY M. S., BROWN M. L., WHITE J. L. CHROMATOGRAPHICALLY HOMOGENEOUS LECITHIN FROM EGG PHOSPHOLIPIDS. J Am Oil Chem Soc. 1965 Jan;42:53–56. doi: 10.1007/BF02558256. [DOI] [PubMed] [Google Scholar]
  15. Schwarz G., Gerke H., Rizzo V., Stankowski S. Incorporation kinetics in a membrane, studied with the pore-forming peptide alamethicin. Biophys J. 1987 Nov;52(5):685–692. doi: 10.1016/S0006-3495(87)83263-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Schwarz G., Savko P. Structural and dipolar properties of the voltage-dependent pore former alamethicin in octanol/dioxane. Biophys J. 1982 Aug;39(2):211–219. doi: 10.1016/S0006-3495(82)84510-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schwarz G., Stankowski S., Rizzo V. Thermodynamic analysis of incorporation and aggregation in a membrane: application to the pore-forming peptide alamethicin. Biochim Biophys Acta. 1986 Sep 25;861(1):141–151. doi: 10.1016/0005-2736(86)90573-0. [DOI] [PubMed] [Google Scholar]
  18. Todd A. P., Mehlhorn R. J., Macey R. I. Amine spin probe permeability in sonicated liposomes. J Membr Biol. 1989 Jul;109(1):53–64. doi: 10.1007/BF01870790. [DOI] [PubMed] [Google Scholar]
  19. Vogel H. Comparison of the conformation and orientation of alamethicin and melittin in lipid membranes. Biochemistry. 1987 Jul 14;26(14):4562–4572. doi: 10.1021/bi00388a060. [DOI] [PubMed] [Google Scholar]
  20. Wille B., Franz B., Jung G. Location and dynamics of alamethicin in unilamellar vesicles and thylakoids as model systems. A spin label study. Biochim Biophys Acta. 1989 Nov 17;986(1):47–60. doi: 10.1016/0005-2736(89)90271-x. [DOI] [PubMed] [Google Scholar]
  21. Woolley G. A., Deber C. M. A lipid vesicle system for probing voltage-dependent peptide-lipid interactions: application to alamethicin channel formation. Biopolymers. 1989 Jan;28(1):267–272. doi: 10.1002/bip.360280127. [DOI] [PubMed] [Google Scholar]
  22. Yantorno R., Takashima S., Mueller P. Dipole moment of alamethicin as related to voltage-dependent conductance in lipid bilayers. Biophys J. 1982 May;38(2):105–110. doi: 10.1016/S0006-3495(82)84536-0. [DOI] [PMC free article] [PubMed] [Google Scholar]

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