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.
Similar content being viewed by others
References
Allen R, Melchionna S, Hansen J-P (2002) Intermittent permeation of cylindrical nanopores by water. Phys Rev Lett 89:175502–175504
Archer DB (1976) Effect of the lipid composition of Mycoplasma mycoides subspecies capri and phosphatidyl choline vesicles upon the action of polyene antibiotics. Biochim Biophys Acta 436:68–76
Barenholz Y (2002) Cholesterol and other membrane active sterols: from membrane evolution to “rafts”. Prog Lipid Res 41:1–5
Beckstein O, Sansom MSP (2004) The influence of geometry, surface character and flexibility on the permeation of ions and water though biological pores. Phys Biol 1:42–52
Boda D, Valisko M, Eisenberg B, Nonner W, Henderson D, Gillespie D (2007) Combined effects of pore radius and protein dielectric coefficient on the selectivity of a calcium channel. Phys Rev Lett 98:168102–168104
Bolard J, Legrand P, Heitz F, Cybulska B (1991) One-sided action of amphotericin B on cholesterol-containing membranes is determined by its self-association in the medium. Biochemistry 30:5707–5715
Cass A, Finkelstein A, Krespi V (1970) The ion permeability induced in thin lipid membranes by the polyene antibiotics nystatin and amphotericin B. J Gen Physiol 56:100–124
Ciana A, Balduini C, Minetti G (2005) Detergent-resistance membranes in human erythrocytes and their connection to the membrane skeleton. J Biosci 30:317–328
Cohen BE (1975) The permeability of liposomes to non-electrolytes. II. The effect of nystatin and gramicidin A. J Membr Biol 20:235–260
Cohen BE (1992) A sequential mechanism for the formation of aqueous channels by amphotericin B in liposomes. The effect of sterols and phospholipid composition. Biochim Biophys Acta 1108:49–58
Cohen BE (1998) Amphotericin B toxicity and lethality: a tale of two channels. Int J Pharmaceutics 162:95–106
Coutinho A, Silva L, Fedorov A, Prieto M (2004) Cholesterol and ergosterol influence nystatin surface aggregation: relation to pore formation. Biophys J 87:3264–3276
Crepaldi Domingues C, Ciana A, Buttafava A, Balduini C, De Paula E, Minetti G (2009) Resistance of human erythrocyte membranes to triton X-100 and C12E6. J Membr Biol 227:39–48
Cybulska B, Herve M, Borowski E, Gary-Bobo CM (1986) Effect of the polar head structure of polyene macrolide antifungal antibiotics on the mode of permeabilization of ergosterol- and cholesterol-containing lipidic vesicles studied by 31P-NMR. Mol Pharmacol 29:293–298
Cybulska B, Bolard J, Seksek O, Czerwinski A, Borowski E (1995) Identification of the structural elements of amphotericin B and other macrolide antibiotics of the heptaene group influencing the ionic selectivity of the permeability pathways formed in the red cell membrane. Biochim Biophys Acta 1240:167–178
Czub J, Baginski M (2006) Comparative molecular dynamics study of lipid membranes containing cholesterol and ergosterol. Biophys J 90:2368–2382
De Kruijff B, Demel RA (1974) Polyene antibiotic–sterol interactions in membranes of Acholesplasma laidlawii cells and lecithin liposomes. III. Molecular structure of the polyene antibiotic-cholesterol complexes. Biochim Biophys Acta 339:57–70
De Kruijff B, Gerritsen WJ, Oerlemanns A, Van Dick PW, Demel RA, Van Deenen LLM (1974) Polyene antibiotic–sterol interactions in membranes of Acholeplasma laidlawii cell and lecithin liposomes. II. Temperature dependence of the polyene antibiotic–sterol complex formation. Biochim Biophys Acta 339:44–56
Deuticke B, Zollner C (1972) Lack of influence of membrane cholesterol on anion and nonelectrolyte permeability of pig erythrocytes. Biochim Biophys Acta 266:726–731
Deuticke B, Kim M, Zollner C (1973) The influence of amphotericin B on the permeability of mammalian erythrocytes to nonelectrolytes, anions and cations. Biochim Biophys Acta 318:345–359
Dodge JT, Mitchell C, Hanahan DJ (1963) The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochim Biophys 100:119–130
Drabkin DL, Austin JH (1932) Spectroscopy studies. I. Spectrophotometric constants for common hemoglobin derivatives in human, dog and rabbit blood. J Biol Chem 98:719–733
Ernst C, Lematre J, Tinnert H, Dupont G, Grange J (1979) Interaction between an antifungal heptaene, amphotericin B and cholesterol in vitro, as detected by circular dichroism and absorption. Influence of temperature. C R Seances Acad Sci D 289:1145–1149
Finkelstein A, Holz R (1973) Aqueous pores created in thin lipid membranes by the polyene antibiotic nystatin and amphotericin B. In: Eisenman G (ed) Membranes, vol 2. Marcel Deckker, New York, pp 377–408
Galla H-J, Luisfetti J (1980) Lateral and transversal diffusion and phase transitions in erythrocyte membranes. An excimer fluorescence study. Biochim Biophys Acta 596:106–117
Gallis HA, Drew RH, Pickard WW (1990) Amphotericin B: 30 years of clinical experience. Rev Infect Dis 12:308–329
Grzybeck M, Chorzlska A, Bok E, Hryniewicz A, Czogalla A, Diakowski W, Sikorski AF (2006) Spectrin–phospholipid interactions. Existence of multiple kinds of binding sites? Chem Phys Lipids 141:133–141
Haest CWM, Plasa G, Kamp D, Deuticke B (1978) Spectrin as a stabilizer of the phospholipid asymmetry in the human erythrocyte membrane. Biochim Biophys Acta 509:21–32
Heinemann V, Bosse D, Jehn U, Kahny B, Wachholz K, Debus A, Scholz P, Kolb H-J, Wilmanns W (1997) Pharmacokinetics of liposomal amphotericin B (AmBisome) in critically ill patients. Antimicrob Agents Chemother 41:1275–1280
Hladky SB, Rink TJ (1976) Potential difference and the distribution of ions across the human red blood cell membrane: a study of the mechanism by which the fluorescent cation, diS-C3-(5) reports membrane potential. J Physiol 263:287–319
Koumanov KS, Tessier C, Monchilova AB, Rainteau D, Wolf C, Quinn PJ (2005) Comparative lipid analysis and structure of detergent-resistant membrane raft fractions isolated from human and ruminant erythrocytes. Arch Biochim Biophys 434:150–158
Labonia WD, Morelli OH Jr, Gimenez MI, Freuler PV, Morelli OH (1987) Effects of l-carnitine on sodium transport in erythrocytes from dialyzed uremic patients. Kidney Int 32:754–759
Laemmli WK (1970) Cleavage of structural proteins during the assembly of the head by bacteriophage T4. Nature 227:680–685
Lambing HE, Wolf WD, Hartsel SC (1993) Temperature effects on the aggregation state and activity of amphotericin B. Biochim Biophys Acta 1152:185–188
Lange Y, Ye J, Steck TL (2007) Scrambling of phospholipids activates red cell membrane cholesterol. Biochemistry 46:2233–2238
Legrand P, Romero EA, Cohen BE, Bolard J (1992) Effects of aggregation and solvent on the toxicity of amphotericin B to human erythrocytes. Antimicrob Agents Chemother 36:2518–2522
Nebl T, Pestonjamasp KN, Leszyk JD, Crowley JL, Oh SW, Luna EJ (2002) Proteomic analysis of a detergent-resistant membrane skeleton from neutrophil plasma membranes. J Biol Chem 277:43399–43409
Ohvo-Rekila H, Ramstedt B, Lppimaki P, Slotte JP (2002) Cholesterol interactions with phospholipids in membranes. Prog Lipid Res 41:66–97
Ramos H, Attias de Murciano A, Cohen BE, Bolard J (1989) The polyene antibiotic amphotericin B acts as a Ca2+ ionophore across sterol-containing liposomes. Biochim Biophys Acta 982:303–306
Ramos H, Valdivieso E, Gamargo M, Dagger F, Cohen BE (1996) Amphotericin B kills unicellular leishmanias by forming aqueous pores permeable to small cations and anions. J Membr Biol 152:65–75
Readio J, Bittman R (1972) Equilibrium binding of amphotericin B and its methyl ester and borate complex to sterols. Biochim Biophys Acta 685:219–224
Rogers PD, Kramer RE, Chapman SW, Cleary JD (1999) Amphotericin B-induced interleukin-1beta expression in human monocyte cells is calcium and calmodulin dependent. J Infect Dis 180:1259–1266
Romero PJ, Romero EA (1997) Differences in Ca2+ pumping activity between sub-populations of human red cells. Cell Calcium 21:353–358
Salzer U, Prohaska R (2001) Stomatin, flotillin-1, and flotillin-2 are major integral proteins of erythrocyte lipid rafts. Blood 97:1141–1143
Shvinka N, Caffier G (1994) Cation conductance and efflux induced by polyene antibiotics in the membrane of skeletal muscle fiber. Biophys J 67:143–152
Silva L, Coutinho A, Fedorova A, Prieto M (2006) Competitive binding of cholesterol and ergosterol to the polyene antibiotic nystatin. A fluorescent study. Biophys J 90:3625–3631
Simons K, Vaz WL (2004) Model systems, lipid rafts and cell membranes. Annu Rev Biophys Biomol Struc 33:269–295
Sundar S, Chakravarty J, Rai VK, Agrawal N, Singh SP, Chauhan V, Murray HW (2007) Amphotericin B treatment for Indian visceral leishmaniasis: response to 15 daily versus alternate-day infusions. Clin Infect Dis 45:556–561
Szponarski W, Bolard J (1987) Temperature-dependent modes for the binding of the polyene amphotericin B to human erythrocyte membranes: a circular dichroism study. Biochim Biophys Acta 897:229–237
Tanaka KI, Ohnishi S (1976) Heterogeneity in the fluidity of intact erythrocyte membrane and its homogenization upon hemolysis. Biochim Biophys Acta 426:218–231
Van Hoogevest P, DeKruijff B (1978) Effect of amphotericin B on cholesterol-containing liposomes of egg phosphatidylcholine and didocosenoyl phosphatidylcholine. A refinement of the model for the formation of pores by amphotericin B in membranes. Biochim Biophys Acta 511:397–407
Venegas V, Gonzalez-Damian J, Celis H, Ortega-Blake I (2003) Amphotericin B channels in the bacterial membrane: role of sterol and temperature. Biophys J 85:2323–2332
Vertut-Doi A, Hannaert P, Bolard J (1988) The polyene antibiotic amphotericin B inhibits the Na+/K+ pump of human erythrocytes. Biochem Biophys Res Commun 157:692–697
Wietzerbin J, Szponarski W, Gary-Bobo C (1990) Kinetic study of interaction between [14C] amphotericin B derivatives and human erythrocytes; relationship between binding and induced K+ leak. Biochim Biophys Acta 1026:93–98
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
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
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00232-009-9187-z