Multiple ion binding sites in I h channels of rod photoreceptors from tiger salamanders

Publisher Website
Google Scholar
Abstract
The mechanism of ion permeation in K+/Na+-permeable I h channels of tiger salamander rod photoreceptors was investigated using the whole-cell voltage-clamp technique. I h channels showed features indicative of pores with multiple ion binding sites: in mixtures of K+ and thallium Tl+, the amplitude of the time-dependent current showed an anomalous mole fraction dependence, and K+ permeation was blocked by other permeant ions (with K 0.5 values: Tl+, 44 μM; Rb+, 220 μM and NH4 +, 1100 /gmM) as well as by essentially impermeant ions (Cs+, 22 μM Ba2+, 9200 μM) which apparently block I h by binding in the pore. In contrast, Na+ had little blocking action on K+ permeation. The block by all of these ions was sensitive to external K+ with the block by Cs+ being the least sensitive. Na+ was more effective than K+ in reducing the block by Tl+, Rb+ and NH4 +, but was less effective for the block by Cs+ and Ba2+. The blocking action of Cs+ and Ba2+ was non-competitive, suggesting that they block I h channels at independent sites. Based on the efficacy of block by the different ions, the degree to which K+ and Na+ antagonize this block and the noncompetitive blocking action of Cs+ and Ba2+, the permeation pathway of I h channels appears to contain at least three ion binding sites with at least two sites having a higher affinity for K+ over Na+ and another site with a higher affinity for Na+ over K+.