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The Journal of General Physiology, Vol 53, 133-156, Copyright © 1969 by The Rockefeller University Press

ARTICLE

The Effect of Amphotericin B on the Water and Nonelectrolyte Permeability of Thin Lipid Membranes

Thomas E. Andreoli 1, Vincent W. Dennis 1, and Ann M. Weigl 1

1 From the Department of Physiology and Pharmacology (Division of Clinical Physiology) and the Department of Medicine (Division of Nephrology), Duke University Medical Center, Durham, North Carolina 27706

This paper reports the effects of amphotericin B, a polyene antibiotic, on the water and nonelectrolyte permeability of optically black, thin lipid membranes formed from sheep red blood cell lipids dissolved in decane. The permeability coefficients for the diffusion of water and nonelectrolytes (PDDi) were estimated from unidirectional tracer fluxes when net water flow (Jw) was zero. Alternatively, an osmotic water permeability coefficient (Pf) was computed from Jw when the two aqueous phases contained unequal solute concentrations. In the absence of amphotericin B, when the membrane solutions contained equimolar amounts of cholesterol and phospholipid, Pf was 22.9 ± 4.6 µsec-1 and PDDHDH2O was 10.8 ± 2.4 µsec-1. Furthermore, PDDi was < 0.05 µsec-1 for urea, glycerol, ribose, arabinose, glucose, and sucrose, and sigmai, the reflection coefficient of each of these solutes was one. When amphotericin B (10-6 M) was present in the aqueous phases and the membrane solutions contained equimolar amounts of cholesterol and phospholipid, PDDHDH2O was 18.1 ± 2.4 µsec-1; Pf was 549 ± 143 µsec-1 when glucose, sucrose, and raffinose were the aqueous solutes. Concomitantly, PDDi varied inversely, and sigmai directly, with the effective hydrodynamic radii of the solutes tested. These polyene-dependent phenomena required the presence of cholesterol in the membrane solutions. These data were analyzed in terms of restricted diffusion and filtration through uniform right circular cylinders, and were compatible with the hypothesis that the interactions of amphotericin B with membrane-bound cholesterol result in the formation of pores whose equivalent radii are in the range 7 to 10.5 A.

Submitted on August 27, 1968


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