divalent ion trapping inside potassium channels of human t.pdfVIP

Divalent Ion Trapping Inside Potassium Channels of Human T Lymphocytes S. GRISSMER a n d M. D. CAHALAN From the Department of Physiology and Biophysics, University of California, Irvine, Califor- nia 92717 ABSTRACT Using the patch-clamp whole-cell recording technique, we investi- gated the influence of external Ca 2+, Ba~+, K +, Rb +, and internal Ca2+ on the rate of K + channel inactivation in the human T lymphocyte-derived cell line, Jurkat E6-1. Raising external Ca 2+ or Ba~+, or reducing external K +, accelerated the rate of the K + current decay during a depolarizing voltage pulse. External Ba~+ also produced a use-dependent block of the K + channels by entering the open channel and becoming trapped inside. Raising internal Ca 2+ accelerated inactivation at lower concentrations than external Ca ~+, but increasing the Ca z+ buffering with BAPTA did not affect inactivation. Raising [K+]o or adding Rb + slowed inactiva- tion by competing with divalent ions. External Rb + also produced a use-dependent removal of block of K + channels loaded with Ba2+ or Ca 2+. From the removal of this block we found that under normal conditions ~25% of the channels were loaded with Ca~+, whereas under conditions with 10 #M internal Ca ~+ the propor- tion of channels loaded with Ca ~+ increased to ~50%. Removing all the divalent cations from the external and internal solution resulted in the induction of a non- selective, voltage-independent conductance. We conclude that Ca 2+ ions from the outside or the in