Direct block of cloned hKv1.5 channel by cytochalasins, actin-disrupting agents.
The action of cytochalasins, actin-disrupting agents on human Kv1.5 channel (hKv1.5) stably expressed in Ltk(-) cells was investigated using the whole cell patch-clamp technique. Cytochalasin B inhibited hKv1.5 currents rapidly and reversibly at +60 mV in a concentration-dependent manner with an IC(50) of 4.2 microM. Cytochalasin A, which has a structure very similar to cytochalasin B, inhibited hKv1.5 (IC(50) of 1.4 microM at +60 mV). Pretreatment with other actin filament disruptors cytochalasin D and cytochalasin J, and an actin filament stabilizing agent phalloidin had no effect on the cytochalasin B-induced inhibition of hKv1.5 currents. Cytochalasin B accelerated the decay rate of inactivation for the hKv1.5 currents. Cytochalasin B-induced inhibition of the hKv1.5 channels was voltage dependent with a steep increase over the voltage range of the channel's opening. However, the inhibition exhibited voltage independence over the voltage range in which channels are fully activated. Cytochalasin B produced no significant effect on the steady-state activation or inactivation curves. The rate constants for association and dissociation of cytochalasin B were 3.7 microM/s and 7.5 s(-1), respectively. Cytochalasin B produced a use-dependent inhibition of hKv1.5 current that was consistent with the slow recovery from inactivation in the presence of the drug. Cytochalasin B (10 microM) also inhibited an ultrarapid delayed rectifier K(+) current (I(K,ur)) in human atrial myocytes. These results indicate that cytochalasin B primarily blocks activated hKv1.5 channels and endogenous I(K,ur) in a cytoskeleton-independent manner as an open-channel blocker.
Choi
Bok Hee
BH
Department of Pharmacology, Chonbuk National University Medical School, Chonju, Chonbuk 561-180, Republic of Korea.
Park
Jung-Ah
JA
Kim
Kyung-Ryoul
KR
Lee
Ggot-Im
GI
Lee
Yong-Tae
YT
Choe
Huhn
H
Ko
Seong-Hoon
SH
Kim
Min-Ho
MH
Seo
Yeon-Ho
YH
Kwak
Yong-Geun
YG
D000199:Actins / Q000187:drug effects*; D000818:Animals; D003572:Cytochalasins / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D051666:Kv1.5 Potassium Channel; D008564:Membrane Potentials / Q000187:drug effects; D051379:Mice; D032383:Myocytes, Cardiac / Q000187:drug effects / Q000378:metabolism; D018408:Patch-Clamp Techniques; D024642:Potassium Channels, Voltage-Gated / Q000187:drug effects* / Q000378:metabolism; D013997:Time Factors; D014162:Transfection
D000199:Actins; D003572:Cytochalasins; C496981:KCNA5 protein, human; C496982:Kcna5 protein, mouse; D051666:Kv1.5 Potassium Channel; D024642:Potassium Channels, Voltage-Gated