Mutations in Na1.5 Reveal Calcium-Calmodulin Regulation of Sodium Channel

Overview

Journal Front Physiol

Date 2019 Jun 25

PMID 31231243

Citations 8

Authors

Eyal Nof

Leonid Vysochek

Eshcar Meisel

Elena Burashnikov

Charles Antzelevitch

Jerome Clatot

Roy Beinart

David Luria

Michael Glikson

Shimrit Oz

Affiliations

Soon will be listed here.

Abstract

Mutations in the gene, encoding the cardiac voltage-gated sodium channel Na1.5, are associated with inherited cardiac arrhythmia and conduction disease. Ca-dependent mechanisms and the involvement of β-subunit (Naβ) in Na1.5 regulation are not fully understood. A patient with severe sinus-bradycardia and cardiac conduction-disease was genetically evaluated and compound heterozygosity in the gene was found. Mutations were identified in the cytoplasmic DIII-IV linker (K1493del) and the C-terminus (A1924T) of Na1.5, both are putative CaM-binding domains. These mutants were functionally studied in human embryonic kidney (HEK) cells and HL-1 cells using whole-cell patch clamp technique. Calmodulin (CaM) interaction and cell-surface expression of heterologously expressed Na1.5 mutants were studied by pull-down and biotinylation assays. The mutation K1493del rendered Na1.5 non-conductive. Na1.5 altered the gating properties of co-expressed functional Na1.5, in a Ca and Naβ1-dependent manner. Na1.5 impaired Naβ1-dependent gating regulation. Ca-dependent CaM-interaction with Na1.5 was blunted in Na1.5. Electrical charge substitution at position 1493 did not affect CaM-interaction and channel functionality. Arrhythmia and conduction-disease -associated mutations revealed Ca-dependent gating regulation of Na1.5 channels. Our results highlight the role of Na1.5 DIII-IV linker in the CaM-binding complex and channel function, and suggest that the Ca-sensing machinery of Na1.5 involves Naβ1.

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