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Title: Use-dependent block of the voltage-gated Na+ channel by tetrodotoxin and saxitoxin: Effect of pore mutations that change ionic selectivity

Journal Article · · Journal of General Physiology
 [1];  [2];  [3]
  1. Vertex Pharmaceuticals Inc., San Diego, CA (United States)
  2. Institut de Pharmacologie et Toxicologie de l’Université, Lausanne (Switzerland)
  3. Sandia National Lab. (SNL-NM), Albuquerque, NM (United States). Nanobiology Dept.; Univ. of New Mexico, Albuquerque, NM (United States). School of Medicine. Dept. of Biochemistry and Molecular Biology
+ Show Author Affiliations

Voltage-gated Na+ channels (NaV channels) are specifically blocked by guanidinium toxins such as tetrodotoxin (TTX) and saxitoxin (STX) with nanomolar to micromolar affinity depending on key amino acid substitutions in the outer vestibule of the channel that vary with NaV gene isoforms. All NaV channels that have been studied exhibit a use-dependent enhancement of TTX/STX affinity when the channel is stimulated with brief repetitive voltage depolarizations from a hyperpolarized starting voltage. Two models have been proposed to explain the mechanism of TTX/STX use dependence: a conformational mechanism and a trapped ion mechanism. In this study, we used selectivity filter mutations (K1237R, K1237A, and K1237H) of the rat muscle NaV1.4 channel that are known to alter ionic selectivity and Ca2+ permeability to test the trapped ion mechanism, which attributes use-dependent enhancement of toxin affinity to electrostatic repulsion between the bound toxin and Ca2+ or Na+ ions trapped inside the channel vestibule in the closed state. Our results indicate that TTX/STX use dependence is not relieved by mutations that enhance Ca2+ permeability, suggesting that ion–toxin repulsion is not the primary factor that determines use dependence. Evidence now favors the idea that TTX/STX use dependence arises from conformational coupling of the voltage sensor domain or domains with residues in the toxin-binding site that are also involved in slow inactivation.

Research Organization:
Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)
Sponsoring Organization:
USDOE National Nuclear Security Administration (NNSA)
Grant/Contract Number:
AC04-94AL85000
OSTI ID:
1625212
Journal Information:
Journal of General Physiology, Vol. 140, Issue 4; ISSN 0022-1295
Publisher:
Rockefeller University PressCopyright Statement
Country of Publication:
United States
Language:
English

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