Spliced isoforms of the cardiac Nav1.5 channel modify channel activation by distinct structural mechanisms

Combining electrophysiology and molecular dynamics simulations, Mancino et al. highlight the role of two alternatively spliced residues in the DI S3-S4 linker that determine channel activation in Nav1.5, and they propose a conserved function in the related isoform Nav1.4. Alternative splicing is an important cellular mechanism that fine tunes the gating properties of both voltage- and ligand-gated ion-channels. The cardiac voltage-gated sodium channel, Nav1.5, is subject to alternative splicing of the DI S3-S4 linker, which generates two types of channels with different activation properties. Here, we show that the gating differences between the adult (mH1) and neonatal (Nav1.5e) isoforms of Nav1.5 are mediated by two amino acid residues: Thr/Ser at position 207 and Asp/Lys at position 211. Electrophysiological experiments, in conjunction with molecular dynamics simulations, revealed that each residue contributes equally to the overall gating shifts in activation, but that the underlying structural mechanisms are different. Asp/Lys at position 211 acts through electrostatic interactions, whereas Thr/Ser at position 207 is predicted to alter the hydrogen bond network at the top of the S3 helix. These distinct structural mechanisms work together to modify movement of the voltage-sensitive S4 helix to bring about channel activation. Interestingly, mutation of the homologous Asp and Thr residues of the skeletal muscle isoform, Nav1.4, to Lys and Ser, respectively, confers a similar gating shift in channel activation, suggesting that these residues may fulfill a conserved role across other Nav channel family members.

Automated summary

Using electrophysiology and molecular dynamics simulations, this study highlights the role of two alternatively spliced residues in the DI S3-S4 linker in channel activation of Nav1.5 and suggests a conserved function in Nav1.4 isoform. Understanding the structural mechanisms of alternative splicing in gating properties of voltage-gated ion channels can provide insights into therapeutic target identification and drug development for related cardiac disorders.