Bilobal architecture is a requirement for calmodulin signaling to Ca_V1.3 channels

Calmodulin (CaM) regulation of voltage-gated calcium (CaV) channels is a powerful Ca²⁺ feedback mechanism that adjusts Ca²⁺ influx, affording rich mechanistic insights into Ca²⁺ decoding. CaM possesses a dual-lobed architecture, a salient feature of the myriad Ca²⁺-sensing proteins, where two homologous lobes that recognize similar targets hint at redundant signaling mechanisms. Here, by tethering CaM lobes, we demonstrate that bilobal architecture is obligatory for signaling to CaV channels. With one lobe bound, CaV carboxy tail rearranges itself, resulting in a preinhibited configuration precluded from Ca²⁺ feedback. Reconstitution of two lobes, even as separate molecules, relieves preinhibition and restores Ca²⁺ feedback. CaV channels thus detect the coincident binding of two Ca²⁺-free lobes to promote channel opening, a molecular implementation of a logical NOR operation that processes spatiotemporal Ca²⁺ signals bifurcated by CaM lobes. Overall, a unified scheme of CaV channel regulation by CaM now emerges, and our findings highlight the versatility of CaM to perform exquisite Ca²⁺ computations.