![Figure 11. Mutation analysis of three residues in the S5 segment predicted to form intrasubunit interactions with the pore helix of Slo2.1. (A) MthK-based homology model of a single Slo2.1 subunit highlighting Phe240 in the pore helix (PH) and three potential interacting residues in the S5 segment. (B) Peak outward ISlo2.1 measured at 0 mV in the absence (control) and presence of 1 and 6 mM NFA for mutant channels with indicated point mutations in S5 (n = 5–7). (C) [NFA]–response relationship for L209T Slo2.1 channels. ISlo2.1 was measured at 0 mV and normalized to the peak current measured in the presence of 3 mM NFA. Data were fitted with a logistic equation (smooth curve). EC50 = 106 ± 9 µM, nH = 1.39 ± 0.12 (n = 6). (D) I-Vt relationships for L209T ISlo2.1 in oocytes bathed in KCM211 and K104 solutions. (E) I-Vt relations for L209T ISlo2.1 before and after treatment with 1 mM NFA (n = 10). (F) I-Vt relationship for L209T ISlo2.1 before and after intracellular NaCl loading. For data presented in C–F, oocytes were injected with 0.7 ng cRNA and currents were recorded 1 d later. Error bars indicate mean ± SEM.](https://cdn.rupress.org/rup/content_public/journal/jgp/142/5/10.1085_jgp.201311064/4/jgp_201311064_fig11.jpeg?Expires=1773587799&Signature=nxsdkeU7qeRV-5xH319T6p3tAn68yChZ6D~8GXa~8HqcC9TmFDP5WRqMVF4yqcRzTceglHBxiOBUjnWIZb1JZxhX~RDLA3TYHQsBBd8WDCrpJcTsbhszZChbKK~dHzneh5CDZxrOw2WfjQEjPW0XZ132Rcrm84IjH-XeLVvAMqF35agmR3oMNy4UY2XBf~iO1n9flpHysDmikprR-wBsMsc2YHkd-8~JLwYmYZHcD3nBAwZ34OETppfucpRap46GkBm1CE5wtIiJC~Jy887dkqQ3UrtaeEfEz6rReAo5RGpUSGTuyJ6il4jPXGN4J1dMnYYDPAfCcrUZELJP958zQQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Mutation analysis of three residues in the S5 segment predicted to form intrasubunit interactions with the pore helix of Slo2.1. (A) MthK-based homology model of a single Slo2.1 subunit highlighting Phe240 in the pore helix (PH) and three potential interacting residues in the S5 segment. (B) Peak outward ISlo2.1 measured at 0 mV in the absence (control) and presence of 1 and 6 mM NFA for mutant channels with indicated point mutations in S5 (n = 5–7). (C) [NFA]–response relationship for L209T Slo2.1 channels. ISlo2.1 was measured at 0 mV and normalized to the peak current measured in the presence of 3 mM NFA. Data were fitted with a logistic equation (smooth curve). EC50 = 106 ± 9 µM, nH = 1.39 ± 0.12 (n = 6). (D) I-Vt relationships for L209T ISlo2.1 in oocytes bathed in KCM211 and K104 solutions. (E) I-Vt relations for L209T ISlo2.1 before and after treatment with 1 mM NFA (n = 10). (F) I-Vt relationship for L209T ISlo2.1 before and after intracellular NaCl loading. For data presented in C–F, oocytes were injected with 0.7 ng cRNA and currents were recorded 1 d later. Error bars indicate mean ± SEM.
