Interconversion of fast and slow gating modes of GCAC1, a Guard Cell Anion Channel

Abstract

For guard-cell protoplasts of Vicia faba L. we elucidated whether the slow (S-type) and rapid (R-type) activating anion channels represent different gating modes of GCAC1, the Guard Cell Anion Channel. In the whole-cell configuration of the patch-clamp technique, GCAC1 was activated in a Ca2+- and nucleotide-dependent manner. Cell-free outside-out membrane patches were isolated to determine the relative contribution of different gating modes or channel types to the overall anion current in this cell type. Within 2–15 min after the loss of cytoplasmic control through patch excision, depolarization-activated 38-pS channels characterized by flickering openings in the millisecond range convert into a channel of similar conductance but prolonged open times (hundreds of milliseconds) and lack of pronounced voltage-dependence. The rapid (R-type) and slow (S-type) gating modes exhibited similar ion selectivity but different susceptibility towards block by the stilbene derivative DNDS (4,4′-dinitrostilbene-2,2′-disulfonic acid). On R-type channels DNDS caused a flickering block and a shift in the threshold potential of activation whereas S-type channels remained unaffected. Because of the striking similarities of both channels with respect to single-channel conductance and relative permeability sequence on the one hand, but time-dependent conversion of R- into S-type gating after patch-excision on the other, we conclude that the mode of action of GCAC1 is under the control of cytoplasmic factors.