Inhibition of the activity of T lymphocyte Kv1.3 channels by extracellular zinc.

The inhibitory effect of zinc on voltage-gated Kv1.3 channels in human T lymphocytes was investigated using the "whole-cell" patch-clamp technique. Application of 10 and 20 microM Zn caused a concentration-dependent shift of activation midpoint of the whole-cell currents from -19.65+/-1.03 mV (mean+/-SE) under control conditions to 9.84+/-0.66 mV upon application of 20 microM Zn. This effect was saturated at zinc concentrations higher than 20 microM. The activation rate was considerably slower, whereas the deactivation rate was not significantly affected by Zn. Inactivation midpoint was shifted from -53.06+/-0.44 mV under control conditions to -36.05+/-0.48 mV in the presence of 100 microM Zn. Inactivation rate was not significantly affected upon Zn treatment. Whole-cell potassium currents were reduced to about 70% of their control values with no clear concentration dependence in the zinc concentration range from 10 to 100 microM. When raising the zinc concentration to levels above 100 microM, a concentration-dependent inhibition of the whole-cell currents appeared additionally to the changes in channel gating. The channels were half-blocked at the zinc concentration of 346+/-40 microM and the Hill slope coefficient was 1.89+/-0.21. The inhibitory effect of zinc was not complete at micromolar concentrations and was saturated at concentrations higher than 1mM. This inhibitory effect was not accompanied by any further modification in the shift of the activation and inactivation midpoints nor by a slowing of the channel activation rate. The inhibitory effect of zinc was significantly diminished in the presence of 150 mM K(+) in the extracellular solution, whereas the zinc-induced shift of the activation threshold and slowing of the activation kinetics remained unchanged when raising extracellular potassium concentration. It is suggested that zinc acts on two independent binding sites on the channels. Binding to one site that is saturated at concentrations higher than 20 microM affects the channel gating. Binding to another site at concentrations higher than 100 microM inhibits the currents without affecting the channel gating.