Contribution of Ion Binding Affinity to Ion Selectivity and Permeation in KcsA, a Model Potassium Channel
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- 作者：M. L. Renart ; E. Montoya ; A. M. Fern谩ndez ; M. L. Molina ; J. A. Poveda ; J. A. Encinar ; J. L. Ayala ; A. V. Ferrer-Montiel ; J. G贸mez ; A. Morales ; J. M. Gonz谩lez Ros
- 刊名：Biochemistry
- 出版年：2012
- 出版时间：May 8, 2012
- 年：2012
- 卷：51
- 期：18
- 页码：3891-3900
- 全文大小：465K
- 年卷期：v.51,no.18(May 8, 2012)
- ISSN：1520-4995
文摘
Ion permeation and selectivity, key features in ion channel function, are believed to arise from a complex ensemble of energetic and kinetic variables. Here we evaluate the contribution of pore cation binding to ion permeation and selectivity features of KcsA, a model potassium channel. For this, we used E71A and M96V KcsA mutants in which the equilibrium between conductive and nonconductive conformations of the channel is differently shifted. E71A KcsA is a noninactivating channel mutant. Binding of K+ to this mutant reveals a single set of low-affinity K+ binding sites, similar to that seen in the binding of K+ to wild-type KcsA that produces a conductive, low-affinity complex. This seems consistent with the observed K+ permeation in E71A. Nonetheless, the E71A mutant retains K+ selectivity, which cannot be explained on the basis of just its low affinity for this ion. At variance, M96V KcsA is a rapidly inactivating mutant that has lost selectivity for K+ and also conducts Na+. Here, low-affinity binding and high-affinity binding of both cations are detected, seemingly in agreement with both being permeating species in this mutant channel. In conclusion, binding of the ion to the channel protein seemingly explains certain gating, ion selectivity, and permeation properties. Ion binding stabilizes greatly the channel and, depending upon ion type and concentration, leads to different conformations and ion binding affinities. High-affinity states guarantee binding of specific ions and mediate ion selectivity but are nonconductive. Conversely, low-affinity states would not discriminate well among different ions but allow permeation to occur.