The pore, not cytoplasmic domains, underlies inactivation in a prokaryotic sodium channel

Evgeny Pavlov, Christopher Bladen, Robert Winkfein, Catherine Diao, Perry Dhaliwal, Robert J. French

Research output: Contribution to journalArticle

Abstract

Kinetics and voltage dependence of inactivation of a prokaryotic voltage-gated sodium channel (NaChBac) were investigated in an effort to understand its molecular mechanism. NaChBac inactivation kinetics show strong, bell-shaped voltage dependence with characteristic time constants ranging from ∼50 ms at depolarized voltages to a maximum of ∼100 s at the inactivation midpoint. Activation and inactivation parameters for four different covalently linked tandem dimer or tandem tetramer constructs were indistinguishable from those of the wild-type channel. Point mutations in the outer part of the pore revealed an important influence of the S195 residue on the process of inactivation. For two mutants (S195D and S195E), the maximal and minimal rates of inactivation observed were increased by ∼2.5-fold, and the midpoint of the steady-state inactivation curve was shifted ∼20 mV in the hyperpolarizing direction, compared to the wild-type channel. Our data suggest that pore vestibule structure is an important determinant of NaChBac inactivation, whereas the inactivation mechanism is independent of the number of free cytoplasmic N- and C-termini in the functional channel. In these respects, NaChBac inactivation resembles C-type or slow inactivation modes observed in other voltage-gated K and Na channels.

Original languageEnglish (US)
Pages (from-to)232-242
Number of pages11
JournalBiophysical Journal
Volume89
Issue number1
DOIs
StatePublished - Jul 2005

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Sodium Channels
Voltage-Gated Sodium Channels
Voltage-Gated Potassium Channels
Point Mutation
Direction compound

ASJC Scopus subject areas

  • Biophysics

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The pore, not cytoplasmic domains, underlies inactivation in a prokaryotic sodium channel. / Pavlov, Evgeny; Bladen, Christopher; Winkfein, Robert; Diao, Catherine; Dhaliwal, Perry; French, Robert J.

In: Biophysical Journal, Vol. 89, No. 1, 07.2005, p. 232-242.

Research output: Contribution to journalArticle

Pavlov, Evgeny ; Bladen, Christopher ; Winkfein, Robert ; Diao, Catherine ; Dhaliwal, Perry ; French, Robert J. / The pore, not cytoplasmic domains, underlies inactivation in a prokaryotic sodium channel. In: Biophysical Journal. 2005 ; Vol. 89, No. 1. pp. 232-242.
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AU - French, Robert J.

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