Reconstructing voltage sensor-pore interaction from a fluorescence scan of a voltage-gated K+ channel.
X-ray crystallography has made considerable recent progress in providing static structures of ion channels. Here we describe a complementary method-systematic fluorescence scanning-that reveals the structural dynamics of a channel. Local protein motion was measured from changes in the fluorescent intensity of a fluorophore attached at one of 37 positions in the pore domain and in the S4 voltage sensor of the Shaker K+ channel. The local rearrangements that accompany activation and slow inactivation were mapped onto the homologous structure of the KcsA channel and onto models of S4. The results place clear constraints on S4 location, voltage-dependent movement, and the mechanism of coupling of S4 motion to the operation of the slow inactivation gate in the pore domain.
Gandhi
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CS
Department of Molecular and Cell Biology, University of California, Berkeley 94720, USA.
Loots
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Isacoff
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EY
D001426:Bacterial Proteins; D005456:Fluorescent Dyes / Q000737:chemistry; D015640:Ion Channel Gating / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015221:Potassium Channels / Q000737:chemistry* / Q000235:genetics*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013050:Spectrometry, Fluorescence / Q000379:methods; D013329:Structure-Activity Relationship
D001426:Bacterial Proteins; D005456:Fluorescent Dyes; D015221:Potassium Channels; C107790:prokaryotic potassium channel