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. 2010 May;135(5):481–494. doi: 10.1085/jgp.201010403

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© 2010 Zhou et al.

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Figure 5. — Replacing the first half of the Slo1 P loop with the Slo3 sequence increases paxilline block affinity and speeds up paxilline block and unblock rates. (A) The time course of onset and recovery of paxilline block for Slo1 (black) and MC8 (red) is shown for representative inside-out patches for each construct. For MC8, 20 nM paxillline was applied (horizontal bar), and 100 nM was used for Slo1. Peak current activated at +200 mV was measured every 2 (Slo1) or 3 (MC8) seconds and plotted against time. (B) Time constants of onset of paxilline block (τ_on) and recovery (τ_off) are plotted for Slo1 (at 100 nM) and MC8 (at 20 nM). Red dots are individual determinations, and bars indicate means and SEM. (C) The forward block rate and paxilline block affinity were calculated for each individual patch for Slo1 and MC8. Block rate (left axis) and affinity (right axis) were determined from τ_on = 1/([PAX]k_f+k_u), k_u = 1/τ_off, and K_b = k_u/k_f.