Optical single-channel analysis of the aerolysin pore in erythrocyte membranes

Scanning microphotolysis (Scamp), a recently developed photobleaching technique, was used to analyze the transport of two small organic anions and one inorganic cation through single pores formed in human erythrocyte membranes by the channel-forming toxin aerolysin secreted by Aeromonas species. The transport rate constants of erythrocyte ghosts carrying a single aerolysin pore were determined to be (1.83 +/- 0.43) x 10(-3) s(-1) for Lucifer yellow, (0.33 +/- 0.10) x 10(-3) s(-1) for carboxyfluorescein, and (8.20 +/- 2.30) x 10(-3) s(-1) for Ca2+. The radius of the aerolysin pore was derived from the rate constants to be 19-23 Angstrom, taking steric hindrance and viscous drag into account. The size of the Ca2+ rate constant implies that at physiological extracellular Ca2+ concentrations (>1 mM) the intracellular Ca2+ concentration would be elevated to the critical level of >1 mu M in much less than a second after formation of a single aerolysin pore in the plasma membrane. Thus changes in the levels of Ca2+ or other critical intracellular components may be more likely to cause cell death than osmotic imbalance.