Biophysical Investigation of Sodium Channel Interaction with β-Subunit Variants Associated with Arrhythmias

Voltage-gated sodium (Na_V) channels help regulate electrical activity of the plasma membrane. Mutations in associated subunits can result in pathological outcomes. Here we examined the interaction of Na_V channels with cardiac arrhythmia-linked mutations in SCN2B and SCN4B, two genes that encode auxiliary β-subunits. To investigate changes in SCN2B ^R137H and SCN4B ^I80T function, we combined three-dimensional X-ray crystallography with electrophysiological measurements on Na_V1.5, the dominant subtype in the heart. SCN4B ^I80T alters channel activity, whereas SCN2B ^R137H does not have an apparent effect. Structurally, the SCN4B ^I80T perturbation alters hydrophobic packing of the subunit with major structural changes and causes a thermal destabilization of the folding. In contrast, SCN2B ^R137H leads to structural changes but overall protein stability is unaffected. SCN4B ^I80T data suggest a functionally important region in the interaction between Na_V1.5 and β4 that, when disrupted, could lead to channel dysfunction. A lack of apparent functional effects of SCN2B ^R137H on Na_V1.5 suggests an alternative working mechanism, possibly through other Na_V channel subtypes present in heart tissue. Indeed, mapping the structural variations of SCN2B ^R137H onto neuronal Na_V channel structures suggests altered interaction patterns.