Role of amino acid residues in transmembrane segments IS6 and IIS6 of the Na+ channel α subunit in voltage-dependent gating and drug block

Vladimir Yarov-Yarovoy, Jancy C. McPhee, Diane Idsvoog, Caroline Pate, Todd Scheuer, William A. Catterall

Research output: Contribution to journalArticle

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Abstract

Alanine-scanning mutagenesis of transmembrane segments IS6 and IIS6 of the rat brain Nav1.2 channel α subunit identified mutations N418A in IS6 and L975A in IIS6 as causing strong positive shifts in the voltage dependence of activation. In contrast, mutations V424A in IS6 and L983A in IIS6 caused strong negative shifts. Most IS6 mutations opposed inactivation from closed states, but most IIS6 mutations favored such inactivation. Mutations L421C and L983A near the intracellular ends of IS6 and IIS6, respectively, exhibited significant sustained Na+ currents at the end of 30-ms depolarizations, indicating a role for these residues in Na+ channel fast inactivation. These residues, in combination with residues at the intracellular end of IVS6, are well situated to form an inactivation gate receptor. Mutation I409A in IS6 reduced the affinity of the local anesthetic etidocaine for the inactivated state by 6-fold, and mutations I409A and N418A reduced use-dependent block by etidocaine. No IS6 or IIS6 mutations studied affected inactivated-state affinity or use-dependent block by the neuroprotective drug sipatrigine (compound 619C89). These results suggest that the local anesthetic receptor site is formed primarily by residues in segments IIIS6 and IVS6 with the contribution of a single amino acid in segment IS6.

Original languageEnglish (US)
Pages (from-to)35393-35401
Number of pages9
JournalJournal of Biological Chemistry
Volume277
Issue number38
DOIs
StatePublished - Sep 20 2002
Externally publishedYes

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Etidocaine
Local Anesthetics
Amino Acids
Mutagenesis
Mutation
Depolarization
Electric potential
Neuroprotective Agents
Alanine
Pharmaceutical Preparations
Rats
Brain
Chemical activation
Scanning
sipatrigine

ASJC Scopus subject areas

  • Biochemistry

Cite this

Role of amino acid residues in transmembrane segments IS6 and IIS6 of the Na+ channel α subunit in voltage-dependent gating and drug block. / Yarov-Yarovoy, Vladimir; McPhee, Jancy C.; Idsvoog, Diane; Pate, Caroline; Scheuer, Todd; Catterall, William A.

In: Journal of Biological Chemistry, Vol. 277, No. 38, 20.09.2002, p. 35393-35401.

Research output: Contribution to journalArticle

Yarov-Yarovoy, Vladimir ; McPhee, Jancy C. ; Idsvoog, Diane ; Pate, Caroline ; Scheuer, Todd ; Catterall, William A. / Role of amino acid residues in transmembrane segments IS6 and IIS6 of the Na+ channel α subunit in voltage-dependent gating and drug block. In: Journal of Biological Chemistry. 2002 ; Vol. 277, No. 38. pp. 35393-35401.
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abstract = "Alanine-scanning mutagenesis of transmembrane segments IS6 and IIS6 of the rat brain Nav1.2 channel α subunit identified mutations N418A in IS6 and L975A in IIS6 as causing strong positive shifts in the voltage dependence of activation. In contrast, mutations V424A in IS6 and L983A in IIS6 caused strong negative shifts. Most IS6 mutations opposed inactivation from closed states, but most IIS6 mutations favored such inactivation. Mutations L421C and L983A near the intracellular ends of IS6 and IIS6, respectively, exhibited significant sustained Na+ currents at the end of 30-ms depolarizations, indicating a role for these residues in Na+ channel fast inactivation. These residues, in combination with residues at the intracellular end of IVS6, are well situated to form an inactivation gate receptor. Mutation I409A in IS6 reduced the affinity of the local anesthetic etidocaine for the inactivated state by 6-fold, and mutations I409A and N418A reduced use-dependent block by etidocaine. No IS6 or IIS6 mutations studied affected inactivated-state affinity or use-dependent block by the neuroprotective drug sipatrigine (compound 619C89). These results suggest that the local anesthetic receptor site is formed primarily by residues in segments IIIS6 and IVS6 with the contribution of a single amino acid in segment IS6.",
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AU - Catterall, William A.

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