Counting Ion and Water Molecules in a Streaming File through the Open-Filter Structure of the K Channel

Abstract

The mechanisms underlying the selective permeation of ions through channel molecules are a fundamental issue related to understanding how neurons exert their functions. The "knock-on" mechanism, in which multiple ions in the selectivity filter are hit by an incoming ion, is one of the leading concepts. This mechanism has been supported by crystallographic studies that demonstrated ion distribution in the structure of the Streptomyces lividans (KcsA) potassium channel. These still pictures under equilibrium conditions, however, do not provide a snapshot of the actual, ongoing permeation processes. To understand the dynamics of permeation, we determined the ratio of the ion and water flow [the water-ion coupling ratio (CR(w-i))] through the KcsA channel by measuring the streaming potential (V(stream)) electrophysiologically. The V(stream) value was converted to the CR(w-i) value, which reveals how individual ion and water molecules are queued in the narrow and short filter during permeation. At high K(+) concentrations, the CR(w-i) value was 1.0, indicating that turnover between the alternating ion and water arrays occurs in a single-file manner. At low K(+), the CR(w-i) value was increased to a point over 2.2, suggesting that the filter contained mostly one ion at a time. These average behaviors of permeation were kinetically analyzed for a more detailed understanding of the permeation process. Here, we envisioned the permeation as queues of ion and water molecules and sequential transitions between different patterns of arrays. Under physiological conditions, we predicted that the knock-on mechanism may not be predominant.