Flourescence measurements reveal stoichiometry of K+ channels formed by modulatory and delayed rectifier α-subunits

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Abstract

Modulatory α-subunits, which comprise one-fourth of all voltage-gated K+ channel (Kv) α-subunits, do not assemble into homomeric channels, but selectively associate with delayed rectifier Kv2 subunits to form heteromeric channels of unknown stoichiometry. Their distinct expression patterns and unique functional properties have made these channels candidate molecular correlates for a broad set of native K+ currents. Here, we combine FRET and electrophysiological measurements to determine the stoichiometry and geometry of heteromeric channels composed of the delayed rectifier Kv2.1 subunit and the modulatory Kv9.3 α-subunit. Kv channel α-subunits were fused with GFP variants, and heteromerization of different combinations of tagged and untagged α-subunits was studied. FRET, evaluated by acceptor photobleaching, was only observed upon formation of functional channels. Our results, obtained from two independent experimental paradigms, suggest the formation of heteromeric Kv2.1/Kv9.3 channels of fixed stoichiometry consisting of three Kv2.1 subunits and one Kv9.3 subunit. Strikingly, despite this uneven stoichiometry, we find that heteromeric Kv2.1 Kv9.3 channels maintain a pseudo-symmetric arrangement of subunits around the central pore.

Original languageEnglish
Pages (from-to)6160-6165
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume102
Issue number17
DOIs
StatePublished - Apr 26 2005
Externally publishedYes

Keywords

  • FRET
  • Potassium channel
  • Silent subunit
  • Voltage-gated K channel

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