Two conformational states involved in the use-dependent TTX blockade of human cardiac Na+ channel.
We used a fast inactivation-deficient mutant (QQQ) of the human heart Na+ channel alpha-subunit (hH1a) to assess the influence of the inactivation gate on tetrodotoxin (TTX) use-dependent block (UDB) and postrepolarization block (PRB). PRB had similar time courses in both channels, suggesting no direct interaction of the inactivation gate with the TTX binding site. The UDB saturated with high concentrations of TTX in hH1a but not in QQQ, revealing the modulatory action of fast inactivation on UDB. TTX did not stabilize the inactivated states of QQQ, and the extra block developing during long depolarizations suggests a higher-affinity site involved in the gating of the channel. These results cannot be solely explained by a slow recovery from the block in the inactivated states. They suggest a common use-dependent block mechanism for hH1a and QQQ involving a high-affinity site. We propose that an activated state is primarily responsible for UDB during short depolarization in the range of the action potential plateau and that fast inactivation modulates the accessibility of the toxin to this site.
Dumaine
R
R
Department of Molecular Physiology, Baylor College of Medicine, Houston, Texas 77030, USA.
Hartmann
H A
HA
D000818:Animals; D004594:Electrophysiology; D006801:Humans; D015640:Ion Channel Gating; D016297:Mutagenesis, Site-Directed; D009206:Myocardium / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism; D026941:Sodium Channel Blockers; D015222:Sodium Channels / Q000235:genetics / Q000502:physiology; D013779:Tetrodotoxin / Q000494:pharmacology*; D014981:Xenopus
D026941:Sodium Channel Blockers; D015222:Sodium Channels; D013779:Tetrodotoxin