SUMOylation of the Cardiac Sodium Channel NaV1.5 Modifies Inward Current and Cardiac Excitability

Abstract

AbstractBackgroundDecreased peak sodium current (INa) and increased late sodium current (INa,L), through the cardiac sodium channel NaV1.5 encoded bySCN5A, cause arrhythmias. Many NaV1.5 post-translational modifications have been reported by us and others. A recent report concluded that acute hypoxia increases INa,Lby increasing a Small Ubiquitin-like MOdifier (SUMOylation) at K442-NaV1.5.ObjectiveTo determine whether and by what mechanisms SUMOylation alters INa, INa,Land cardiac electrophysiology.MethodsSUMOylation of NaV1.5 was detected by immunoprecipitation and immunoblotting. INawas measured by patch clamp with/without SUMO1 overexpression in HEK293 cells expressing wild type (WT) or K442R-NaV1.5 and in neonatal rat cardiac myocytes (NRCMs). SUMOylation effects were studiedin vivoby electrocardiograms and ambulatory telemetry using Scn5a heterozygous knockout (SCN5A+/-) mice and the de-SUMOylating protein SENP2 (AAV9-SENP2) or the SUMOylation inhibitor anacardic acid. NaV1.5 trafficking was detected by immunofluorescence.ResultsNaV1.5 was SUMOylated in HEK293 cells, NRCMs and human heart tissue. HyperSUMOylation at NaV1.5-K442 increased INain NRCMs and in HEK cells overexpressing WT but not K442R-Nav1.5. SUMOylation did not alter other channel properties including INa,L. AAV9-SENP2 or anacardic acid treatment of SCN5A+/-mice decreased INa, prolonged QRS duration, and produced heart block and ventricular arrhythmias. SUMO1 overexpression enhanced membrane localization of NaV1.5.ConclusionSUMOylation of K442-Nav1.5 increases peak INawithout changing INa,L, at least in part by altering membrane abundance. Our findings do not support SUMOylation as a mechanism for changes in INa,L. Nav1.5 SUMOylation may modify arrhythmic risk in disease states and represents a potential target for pharmacological manipulation.