Discovery of a functional EF-hand domain within the cardiac sodium channel c-terminus reveals a novel mechanism of direct intracellular calcium regulation

Sodium channels initiate the electrical cascade responsible for cardiac rhythm, and certain life-threatening arrhythmias arise from Na+ channel dysfunction. We propose a novel mechanism for modulation of Na + channel function whereby calcium ions bind directly to the human cardiac Na+ channel (hH1) via an EF-hand motif in the C-terminal domain. A functional role for Ca2+ binding was identified electrophysiologically, by measuring Ca2+-induced modulation of hH1. A small hH1 fragment containing the EF-hand motif was shown to form a structured domain and to bind Ca2+ with affinity characteristic of calcium sensor proteins. Mutations in this domain reduce Ca2+ affinity in vitro and the inactivation gating effects of Ca2+ in electrophysiology experiments. These studies reveal the molecular basis for certain forms of long QT syndrome and other arrhythmia-producing syndromes, and suggest a potential pharmacological target for antiarrhythmic drug design.