Structural basis for electromechanical coupling and pore block of human Cav1.4
摘要
Cav1.4 is an L-type calcium channel (LTCC) essential for visual signal transmission. Here, we present cryo-EM structures of human Cav1.4 alone and in complex with a potent antagonist SR33805, both at resolutions of ~2.9 Å. The fishhook-shaped SR33805 molecule, an indole analogue, penetrates the III-IV fenestration and projects to the central cavity, immediately explaining its mechanism of action. In both structures, VSDII and VSDIII display the completely and partially down conformations, respectively, while the other two VSDs are up. The pore domain exhibits a tightly closed conformation. To probe the molecular determinants for the different conformations of VSDII among the four LTCCs, we solved the structure of human Cav1.1, which features four up VSDs, and generated a Cav1.4 chimera with VSDII replaced by that from Cav1.1. The structure of Cav1.4 chimera shows an up VSDII, suggesting that the conformational determinants may exist within the domain. Consistent with this analysis, substitution of three varied residues resulted in mixed up and down conformations of VSDII in the cryo-EM analysis. Our studies complete the structural gallery of all ten human Cav channels, providing a comprehensive framework for comparative analysis of their working and disease mechanisms and for structure-guided drug discovery.
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