title: Permeability increase induced by Escherichia coli hemolysin A in human macrophages is due to the formation of ionic pores: a patch clamp characterization.
abstract: Escherichia coli hemolysin is known to cause hemolysis of red blood cells by forming hydrophilic pores in their cell membrane. Hemolysin-induced pores have been directly visualized in model systems such as planar lipid membranes and unilamellar vesicles. However this hemolysin, like all the members of a related family of toxins called Repeat Toxins, is a potent leukotoxin. To investigate whether the formation of channels is involved also in its leukotoxic activity, we used patch-clamped human macrophages as targets. Indeed, when exposed to the hemolysin, these cells developed additional pores into their membrane. Such exogenous pores had properties very different from the endogenous channels already present in the cell membrane (primarily K+ channels), but very similar to the pores formed by the toxin in purely lipidic model membranes. Observed properties were: large single channel conductance, cation over anion selectivity but weak discrimination among different cations, quasilinear current-voltage characteristic and the existence of a flickering pre-open state of small conductance. The selectivity properties of the toxin channels appearing in phospholipid vesicles were also investigated, using a specially adapted polarization/depolarization assay, and were found to be completely consistent with that of the current fluctuations observed in excised macrophage patches.
authors.affiliation: Centro CNR-ITC Fisica Stati Aggregati, Povo, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Menestrina
authors.forename: C
authors.initials: C
authors.lastname: Pederzolli
authors.forename: M
authors.initials: M
authors.lastname: Dalla Serra
authors.forename: M
authors.initials: M
authors.lastname: Bregante
authors.forename: F
authors.initials: F
authors.lastname: Gambale
mesh_terms: D001426:Bacterial Proteins / Q000494:pharmacology*; D002460:Cell Line; D002463:Cell Membrane Permeability / Q000187:drug effects*; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins / Q000494:pharmacology*; D006801:Humans; D008055:Lipids; D008264:Macrophages / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D008564:Membrane Potentials / Q000187:drug effects; D009000:Monocytes / Q000166:cytology / Q000187:drug effects; D018408:Patch-Clamp Techniques
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D006460:Hemolysin Proteins; C036507:Hlya protein, E coli; D008055:Lipids