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Formation of Two Different Types of Ion Channels by Amphotericin B in Human Erythrocyte Membranes

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Abstract

The polyene antibiotic amphotericin B (AmB) is known to form aqueous pores in lipid membranes and biological membranes. Here, membrane potential and ion permeability measurements were used to demonstrate that AmB can form two types of selective ion channels in human erythrocytes, differing in their interaction with cholesterol. We show that AmB induced a cation efflux (negative membrane polarization) across cholesterol-containing liposomes and erythrocytes at low concentrations (≤1.0 × 10−6 M), but a sharp reversal of such polarization was observed at concentrations greater than 1.0 × 10−6 M AmB, an indication that aqueous pores are formed. Cation-selective AmB channels are also formed across sterol-free liposomes, but aqueous pores are only formed at AmB concentrations 10 times greater. The effect of temperature on the AmB-mediated K+ efflux across erythrocytes revealed that the energies of activation for channel formation are negative and positive at AmB concentrations that lead predominantly to the formation of cation-selective channels and aqueous pores, respectively. These findings support the conclusion that the two types of AmB channels formed in human erythrocytes differ in their interactions with cholesterol and other membrane components. In effect, a membrane lipid reorganization, as induced by incubation of erythrocytes with tetrathionate, a cross-linking agent of the lipid raft–associated protein spectrin, led to differential changes in the activation parameters for the formation of both types of channels, reflecting the different lipid environments in which such structures are formed.

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Acknowledgement

The authors are indebted to Prof. Pedro J. Romero for providing access to the facilities and equipment of his laboratory at the Central University of Venezuela to perform the experiments with human erythrocytes.

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Authors and Affiliations

  1. Institute of Experimental Biology, Faculty of Sciences, Central University of Venezuela, Caracas, Venezuela

    Eneida A. Romero & Elizabeth Valdivieso

  2. Division of External Activities, National Institute of Allergy and Infectious Diseases, 6700B Rockledge Drive, Bethesda, MD, 20982, USA

    B. Eleazar Cohen

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  1. Eneida A. Romero
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  2. Elizabeth Valdivieso
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  3. B. Eleazar Cohen
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Correspondence to B. Eleazar Cohen.

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Romero, E.A., Valdivieso, E. & Cohen, B.E. Formation of Two Different Types of Ion Channels by Amphotericin B in Human Erythrocyte Membranes. J Membrane Biol 230, 69–81 (2009). https://doi.org/10.1007/s00232-009-9187-z

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  • DOI: https://doi.org/10.1007/s00232-009-9187-z

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