A voltage-sensitive, cation-selective ion channel of Escherichia coli has been reconstituted into liposomes and studied with the patch-clamp method. The single channel conductance was 91 pS in symmetric solutions of 150 mM KCl. Many channels were open most of the time, with frequent brief transitions to closed levels. Multiple conducting units could close and reopen simultaneously, and this apparent cooperativity in gating was increased with depolarizing voltages. Above a voltage threshold, the channels closed irreversibly, often in groups.