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Proc Natl Acad Sci U S A
2013 Mar 26;11013:5217-22. doi: 10.1073/pnas.1219611110.
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State-dependent FRET reports calcium- and voltage-dependent gating-ring motions in BK channels.
Miranda P
,
Contreras JE
,
Plested AJ
,
Sigworth FJ
,
Holmgren M
,
Giraldez T
.
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Large-conductance voltage- and calcium-dependent potassium channels (BK, "Big K+") are important controllers of cell excitability. In the BK channel, a large C-terminal intracellular region containing a "gating-ring" structure has been proposed to transduce Ca(2+) binding into channel opening. Using patch-clamp fluorometry, we have investigated the calcium and voltage dependence of conformational changes of the gating-ring region of BK channels, while simultaneously monitoring channel conductance. Fluorescence resonance energy transfer (FRET) between fluorescent protein inserts indicates that Ca(2+) binding produces structural changes of the gating ring that are much larger than those predicted by current X-ray crystal structures of isolated gating rings.
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