Stabilization of the Conductive Conformation of a Voltage-gated K+ (Kv) Channel

Voltage-gated K + (Kv) channels are molecular switches that sense membrane potential and in response open to allow K + ions to diffuse out of the cell. In these proteins, sensor and pore belong to two distinct structural modules. We previously showed that the pore module alone is a robust yet dynamic structural unit in lipid membranes and that it senses potential and gates open to conduct K + with unchanged fidelity. The implication is that the voltage sensitivity of K + channels is not solely encoded in the sensor. Given that the coupling between sensor and pore remains elusive, we asked whether it is then possible to convert a pore module characterized by brief openings into a conductor with a prolonged lifetime in the open state. The strategy involves selected probes targeted to the filter gate of the channel aiming to modulate the probability of the channel being open assayed by single channel recordings from the sensorless pore module reconstituted in lipid bilayers. Here we show that the premature closing of the pore is bypassed by association of the filter gate with two novel open conformation stabilizers: an antidepressant and a peptide toxin known to act selectively on Kv channels. Such stabilization of the conductive conformation of the channel is faithfully mimicked by the covalent attachment of fluorescein at a cysteine residue selectively introduced near the filter gate. This modulation prolongs the occupancy of permeant ions at the gate. It is this longer embrace between ion and gate that we conjecture underlies the observed stabilization of the conductive conformation. This study provides a new way of thinking about gating. Background: In voltage-gated K + channels, sensor and pore are two structural modules, but their functional coupling remains elusive. Results: Premature pore closing is bypassed by association of the filter gate with novel open conformation stabilizers. Conclusion: Prolonged occupancy of cargo at the gate underlies stabilization. Significance: A gate covered by a lid provides new ways of thinking about gating mechanisms.