Nanoenviroments of the β-Subunit of L-Type Voltage-Gated Calcium Channels in Adult Cardiomyocytes

In cardiomyocytes, Ca²⁺ influx through L-type voltage-gated calcium channels (LTCCs) following membrane depolarization regulates crucial Ca²⁺-dependent processes including duration and amplitude of the action potentials and excitation-contraction coupling. LTCCs are heteromultimeric proteins composed of the Ca_vα₁, Ca_vβ, Ca_vα₂δ and Ca_vγ subunits. Here, using ascorbate peroxidase (APEX2)-mediated proximity labeling and quantitative proteomics, we identified 61 proteins in the nanoenvironments of Ca_vβ₂ in cardiomyocytes. These proteins are involved in diverse cellular functions such as cellular trafficking, cardiac contraction, sarcomere organization and excitation-contraction coupling. Moreover, pull-down assays and co-immunoprecipitation analyses revealed that Ca_vβ₂ interacts with the ryanodine receptor 2 (RyR2) in adult cardiomyocytes, probably coupling LTCCs and the RyR2 into a supramolecular complex at the dyads. This interaction is mediated by the Src-homology 3 domain of Ca_vβ₂ and is necessary for an effective pacing frequency-dependent increase of the Ca²⁺-induced Ca²⁺ release mechanism in cardiomyocytes.