Figure 6. Voltage-dependent rearrangement of the eag domain-CNBHD interaction for zELK channels.(A) Kinetic measurement of Anap fluorescence during a +60 mV depolarization pulse with simultaneous current recordings, in the presence or absence of ATP/Mg2+. (B) and (C) Comparison of the kinetics of the development (B) and the recovery (C) of tmFRET and VDP (n = 4–6). (D) Cartoon illustrating that the VDP of zELK channels involves a rearrangement of the direct interaction between the eag domain and CNBHD. The intersubunit interaction of the eag domain and CNBHD of diagonal subunits is illustrated at hyperpolarizing and depolarizing voltages showing a rearrangement of this interaction accompanies the VDP. Other changes such as pore opening and movement of the S4 are also important for channel activation. The yellow box indicates the intrinsic ligand. PI(4,5)P2 in the inner leaflet of the plasma membrane is shown to highlight its potential role in regulating VDP.DOI: http://dx.doi.org/10.7554/eLife.26355.012Figure 6—figure supplement 1. Measuring the recovery of VDP.(A) Recovery of VDP measured by a voltage protocol with hyperpolarizing recovery pulses of various durations. Representative control current traces without the +60 mV prepulse are shown, and representative current traces with +60 mV prepulse and subsequent 0, 100 and 300 ms recovery pulses are shown. (B) Relationship between the duration of the recovery pulse and V1/2 of zELK channel activation after a +60 mV prepulse (n = 6). Theτ of the exponential fit in red is 136 ms. (C) Relationship between the duration of the recovery pulse and the peak tail-current amplitudes of zELK channels after a +60 mV prepulse (n = 6). Theτ of the exponential fit in red is 293 ms.DOI: http://dx.doi.org/10.7554/eLife.26355.013
Image published in: Dai G and Zagotta WN (2017)
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