Calcium binding and voltage gating in Cx46 hemichannels

The opening of connexin (Cx) hemichannels in the membrane is tightly regulated by calcium (Ca(2+)) and membrane voltage. Electrophysiological and atomic force microscopy experiments indicate that Ca(2+) stabilizes the hemichannel closed state. However, structural data show that Ca(2+) binding induces an electrostatic seal preventing ion transport without significant structural rearrangements. In agreement with the closed-state stabilization hypothesis, we found that the apparent Ca(2+) sensitivity is increased as the voltage is made more negative. Moreover, the voltage and Ca(2+) dependence of the channel kinetics indicate that the voltage sensor movement and Ca(2+) binding are allosterically coupled. An allosteric kinetic model in which the Ca(2+) decreases the energy necessary to deactivate the voltage sensor reproduces the effects of Ca(2+) and voltage in Cx46 hemichannels. In agreement with the model and suggesting a conformational change that narrows the pore, Ca(2+) inhibits the water flux through Cx hemichannels. We conclude that Ca(2+) and voltage act allosterically to stabilize the closed conformation of Cx46 hemichannels.