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pubmed:abstractText |
Permeation, gating, and their interrelationship in an inwardly rectifying potassium (K+) channel, ROMK2, were studied using heterologous expression in Xenopus oocytes. Patch-clamp recordings of single channels were obtained in the cell-attached mode. The gating kinetics of ROMK2 were well described by a model having one open and two closed states. One closed state was short lived (approximately 1 ms) and the other was longer lived (approximately 40 ms) and less frequent (approximately 1%). The long closed state was abolished by EDTA, suggesting that it was due to block by divalent cations. These closures exhibit a biphasic voltage dependence, implying that the divalent blockers can permeate the channel. The short closures had a similar biphasic voltage dependence, suggesting that they could be due to block by monovalent, permeating cations. The rate of entering the short closed state varied with the K+ concentration and was proportional to current amplitude, suggesting that permeating K+ ions may be related to the short closures. To explain the results, we propose a variable intrapore energy well model in which a shallow well may change into a deep one, resulting in a normally permeant K+ ion becoming a blocker of its own channel.
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