Staurosporine directly blocks Kv1.3 channels expressed in Chinese hamster ovary cells.
The effects of staurosporine (ST), a widely used protein kinase C (PKC) inhibitor, were examined on Kv1.3 channels stably expressed in Chinese hamster ovary (CHO) cells using the whole-cell and excised inside-out configurations of the patch clamp technique. In whole-cell recordings, ST, at external concentrations from 300 nM to 10 microM, accelerated the rate of inactivation of Kv1.3 currents and thereby reduced the current at the end of the depolarizing pulse in a concentration-dependent manner with an IC50 of 1.2 microM. The actions of ST were unaffected by pretreatment with another selective PKC inhibitor, chelerythrine, or by including the PKC pseudosubstrate peptide inhibitor, PKC 19-36, in the intracellular solution. Rp-cAMPS, a specific protein kinase A inhibitor, included in intracellular solution did not affect the effects of ST. Furthermore, the same effects of ST on Kv1.3 were also observed in excised inside-out patches when applied to the internal face of the membrane. These effects were completely reversible upon washing. Current-voltage relations for Kv1.3 currents at the end of voltage steps indicated that ST reduced Kv1.3 currents over a wide voltage range. The blockade exhibited a shallow voltage dependence between -10 mV and +40 mV, increasing at more positive potentials. ST had no effect on the voltage dependence of steady-state inactivation. It reduced the tail current amplitude and slowed the deactivation time course, resulting in a crossover phenomenon. These results suggest that the action of ST on Kv1.3 is independent of PKC and PKA inhibition. ST blocks the open state of Kv1.3 channels to produce an apparent acceleration of the inactivation rate.
Choi
J S
JS
Department of Physiology, College of Medicine, The Catholic University of Korea, Seoul.
Hahn
S J
SJ
Rhie
D J
DJ
Jo
Y H
YH
Kim
M S
MS
D000470:Alkaloids; D000818:Animals; D053119:Benzophenanthridines; D016466:CHO Cells; D006224:Cricetinae; D000242:Cyclic AMP / Q000031:analogs & derivatives / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D004305:Dose-Response Relationship, Drug; D004558:Electric Stimulation; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005786:Gene Expression Regulation; D051664:Kv1.3 Potassium Channel; D008564:Membrane Potentials / Q000187:drug effects; D018408:Patch-Clamp Techniques; D010617:Phenanthridines / Q000494:pharmacology; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000235:genetics; D024642:Potassium Channels, Voltage-Gated; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D019311:Staurosporine / Q000494:pharmacology*; D013873:Thionucleotides / Q000494:pharmacology
D000470:Alkaloids; D053119:Benzophenanthridines; D004791:Enzyme Inhibitors; D051664:Kv1.3 Potassium Channel; D010617:Phenanthridines; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D024642:Potassium Channels, Voltage-Gated; D013873:Thionucleotides; C016957:adenosine-3',5'-cyclic phosphorothioate; D000242:Cyclic AMP; C016299:chelerythrine; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D019311:Staurosporine