Pore determinants of KCNQ3 K+ current expression

Frank S. Choveau, Ciria C. Hernandez, Sonya M. Bierbower, Mark S Shapiro

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Abstract

KCNQ3 homomeric channels yield very small macroscopic currents compared with other KCNQ channels or KCNQ2/3 heteromers. Two disparate regions of the channels - the C-terminus and the pore region - have been implicated in governing KCNQ current amplitudes. We previously showed that the C-terminus plays a secondary role compared with the pore region. Here, we confirm the critical role of the pore region in determining KCNQ3 currents. We find that mutations at the 312 position in the pore helix of KCNQ3 (I312E, I312K, and I312R) dramatically decreased KCNQ3 homomeric currents as well as heteromeric KCNQ2/3 currents. Evidence that these mutants were expressed in the heteromers includes shifted TEA sensitivity compared with KCNQ2 homomers. To test for differential membrane protein expression, we performed total internal reflection fluorescence imaging, which revealed only small differences that do not underlie the differences in macroscopic currents. To determine whether this mechanism generalizes to other KCNQ channels, we tested the effects of analogous mutations at the conserved I273 position in KCNQ2, with similar results. Finally, we performed homology modeling of the pore region of wild-type and mutant KCNQ3 channels to investigate the putative structural mechanism mediating these results. The modeling suggests that the lack of current in I312E, I312K, and I312R KCNQ3 channels is due to pore helix-selectivity filter interactions that lock the selectivity filter in a nonconductive conformation.

Original languageEnglish (US)
Pages (from-to)2489-2498
Number of pages10
JournalBiophysical Journal
Volume102
Issue number11
DOIs
StatePublished - Jun 6 2012

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Mutation
Optical Imaging
Membrane Proteins

ASJC Scopus subject areas

  • Biophysics

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Pore determinants of KCNQ3 K+ current expression. / Choveau, Frank S.; Hernandez, Ciria C.; Bierbower, Sonya M.; Shapiro, Mark S.

In: Biophysical Journal, Vol. 102, No. 11, 06.06.2012, p. 2489-2498.

Research output: Contribution to journalArticle

Choveau, FS, Hernandez, CC, Bierbower, SM & Shapiro, MS 2012, 'Pore determinants of KCNQ3 K+ current expression', Biophysical Journal, vol. 102, no. 11, pp. 2489-2498. https://doi.org/10.1016/j.bpj.2012.04.018
Choveau, Frank S. ; Hernandez, Ciria C. ; Bierbower, Sonya M. ; Shapiro, Mark S. / Pore determinants of KCNQ3 K+ current expression. In: Biophysical Journal. 2012 ; Vol. 102, No. 11. pp. 2489-2498.
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