The effects of permeant (K+) ions on polyamine (PA)-induced rectification of cloned strong inwardly rectifying channels (IRK1, Kir2.1) expressed in Xenopus oocytes were examined using patch-clamp techniques. The kinetics of PA-induced rectification depend strongly on external, but not internal K+, concentration. Increasing external [K+] speeds up 'activation' kinetics and shifts rectification to more positive membrane potentials. The shift of rectification is directly proportional to the shift in the K+ reversal potential (E(K)) with slope factors +0.62, +0.81, and +0.91 for 1 mM putrescine (Put), 100μM spermidine and 20 μM spermine (Spm), respectively. The time constant of current activation, resulting from unblock of Spm, also shifts directly in proportion to E(K) with slope factor +1.1. Increasing internal [K+] slows down activation kinetics and has a much weaker relieving effect on block by PA: Spm-induced rectification and time constant of activation (Spm unblock) shift directly in proportion to the corresponding change in E9k) with slope factors -0.15 and +0.31, respectively, for 20 μM Spm. The speed up of activation kinetics caused by increase of external [K+] cannot be reversed by equal increase of internal [K+]. The data are consistent with the hypothesis that the conduction pathway of strong inward rectifiers is a long and narrow pore with multiple binding sites for PA and K+.
- inward rectifier
- potassium channel