Abstract
Sodium channels expressed in oocytes exhibited isoform differences in phasic block by saxitoxin (STX). Neuronal channels (rat IIa co-expressed with beta 1 subunit, Br2a + beta 1) had slower kinetics of phasic block for pulse trains than cardiac channels (RHI). After the membrane was repolarized from a single brief depolarizing step, a test pulse at increasing intervals showed first a decrease in current (post-repolarization block) then eventual recovery in the presence of STX. This block/unblock process for Br2a + beta 1 was 10-fold slower than that for RHI. A model accounting for these results predicts a faster toxin dissociation rate and a slower association rate for the cardiac isoform, and it also predicts a shorter dwell time in a putative high STX affinity conformation for the cardiac isoform. The RHI mutation (Cys374-->Phe), which was previously shown to be neuronal-like with respect to high affinity tonic toxin block, was also neuronal-like with respect to the kinetics of post-repolarization block, suggesting that this single amino acid is important for conferring isoform-specific transition rates determining post-repolarization block. Because the same mutation determines both sensitivity for tonic STX block and the kinetics of phasic STX block, the mechanisms accounting for tonic block and phasic block share the same toxin binding site. We conclude that the residue at position 374, in the putative pore-forming region, confers isoform-specific channel kinetics that underlie phasic toxin block.
Original language | English |
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Pages (from-to) | 1353-1363 |
Number of pages | 11 |
Journal | Biophysical Journal |
Volume | 66 |
Issue number | 5 |
DOIs | |
State | Published - 1994 |
Bibliographical note
Funding Information:We are grateful to Al Goldin for Br2a, Jim Limberis for technical assistance, and Aaron Fox for help with Axobasic 1.0. This work was supported by a grant from the National Institutes of Health (PO1 HL20592) and Grants-in-Aid from the American Heart Association-Metropolitan Chicago (J. Satin, J. W. Kyle). Dr. Makielski is an Established Investigator of the American Heart Association.
ASJC Scopus subject areas
- Biophysics