PT - JOURNAL ARTICLE AU - Aditya Bhattacharji AU - Benjamin Kaplan AU - Thanawath Harris AU - Xiaoguang Qu AU - Markus W. Germann AU - Manuel Covarrubias TI - The Concerted Contribution of the S4-S5 Linker and the S6 Segment to the Modulation of a K<sub>v</sub> Channel by 1-Alkanols AID - 10.1124/mol.106.026187 DP - 2006 Nov 01 TA - Molecular Pharmacology PG - 1542--1554 VI - 70 IP - 5 4099 - http://molpharm.aspetjournals.org/content/70/5/1542.short 4100 - http://molpharm.aspetjournals.org/content/70/5/1542.full SO - Mol Pharmacol2006 Nov 01; 70 AB - Gating of voltage-gated K+ channels (Kv channels) depends on the electromechanical coupling between the voltage sensor and activation gate. The main activation gate of Kv channels involves the COOH-terminal section of the S6 segment (S6-b) and the S4-S5 linker at the intracellular mouth of the pore. In this study, we have expanded our earlier work to probe the concerted contribution of these regions to the putative amphipathic 1-alkanol site in the Shaw2 K+ channel. In the S4-S5 linker, we found a direct energetic correlation between α-helical propensity and the inhibition of the Shaw2 channel by 1-butanol. Spectroscopic structural analyses of the S4-S5 linker supported this correlation. Furthermore, the analysis of chimeric Shaw2 and Kv3.4 channels that exchanged their corresponding S4-S5 linkers showed that the potentiation induced by 1-butanol depends on the combination of a single mutation in the S6 PVPV motif (PVAV) and the presence of the Shaw2 S4-S5 linker. Then, using tandem-heterodimer subunits, we determined that this potentiation also depends on the number of S4-S5 linkers and PVAV mutations in the Kv channel tetramer. Consistent with the critical contribution of the Shaw2 S4-S5 linker, the equivalent PVAV mutation in certain mammalian Kv channels with divergent S4-S5 linkers conferred weak potentiation by 1-butanol. Overall, these results suggest that 1-alkanol action in Shaw2 channels depends on interactions involving the S4-S5 linker and the S6-b segment. Therefore, we propose that amphiphilic general anesthetic agents such as 1-alkanols may modulate gating of the Shaw2 K+ channel by an interaction with its activation gate. The American Society for Pharmacology and Experimental Therapeutics