Voltage-gated sodium channels (Na_v) underlie the rapid upstroke of action potentials in excitable tissues. Binding of channel-interactive proteins is essential for controlling fast and long-term inactivation. In the structure of the complex of the carboxy-terminal portion of Na_v1.5 (CTNa_v1.5) with calmodulin (CaM)–Mg²⁺ reported here, both CaM lobes interact with the CTNa_v1.5. On the basis of the differences between this structure and that of an inactivated complex, we propose that the structure reported here represents a non-inactivated state of the CTNa_v, that is, the state that is poised for activation. Electrophysiological characterization of mutants further supports the importance of the interactions identified in the structure. Isothermal titration calorimetry experiments show that CaM binds to CTNa_v1.5 with high affinity. The results of this study provide unique insights into the physiological activation and the pathophysiology of Na_v channels.