Pore formation by the Escherichia coli hemolysin: evidence for an association-dissociation equilibrium of the pore-forming aggregates.
Lipid bilayer experiments were performed in the presence of hemolysin of Escherichia coli. The toxin had a rather low activity in membranes formed of pure lipids, such as phosphatidylcholine or phosphatidylserine. In membranes from asolectin, a crude lipid mixture from soybean, hemolysin was able to increase the conductance by many orders of magnitude in a steep concentration-dependent fashion, which suggested that several hemolysin molecules could be involved in the conductive unit. Furthermore, the much higher toxin activity in asolectin membranes would be consistent with the assumption that this lipid contains a receptor needed for membrane activity of the toxin. The results of single-channel records showed that the membrane activity of hemolysin is due to the formation of ion-permeable channels with a single-channel conductance of about 500 pS in 0.15 M KCl. The hemolysin channel seemed to be formed by a toxin oligomer which showed an association-dissociation reaction and had a mean lifetime of about 2 s at small transmembrane voltages. The conductance of the hemolysin channels was only moderately dependent on the salt concentration in the aqueous phase. Zero-current membrane potential experiments showed that the hemolysin channel is cation selective. The mobility sequence of the cations in the channel was similar to their mobility sequence in the aqueous phase, which was consistent with the assumption that the hemolysin channel is wide and that the interior field strength is not very high. From the single-channel conductance, a lower limit of about 1.0 nm for the effective channel diameter could be estimated.
Benz
R
R
Lehrstuhl für Biotechnologie, Universität Würzburg, Federal Republic of Germany.
Schmid
A
A
Wagner
W
W
Goebel
W
W
D002414:Cations, Monovalent; D004553:Electric Conductivity; D004926:Escherichia coli / Q000472:pathogenicity*; D006460:Hemolysin Proteins / Q000494:pharmacology*; D066298:In Vitro Techniques; D007473:Ion Channels; D008051:Lipid Bilayers; D046911:Macromolecular Substances; D010743:Phospholipids; D013329:Structure-Activity Relationship; D013499:Surface Properties
D002414:Cations, Monovalent; D006460:Hemolysin Proteins; D007473:Ion Channels; D008051:Lipid Bilayers; D046911:Macromolecular Substances; D010743:Phospholipids