CaV β controls the endocytic turnover of CaV 1.2 L-type calcium channel.

Membrane depolarization activates the multisubunit Ca_V 1.2 L-type calcium channel initiating various excitation coupling responses. Intracellular trafficking into and out of the plasma membrane regulates the channel's surface expression and stability, and thus, the strength of Ca_V 1.2-mediated Ca²⁺ signals. The mechanisms regulating the residency time of the channel at the cell membrane are unclear. Here, we coexpressed the channel core complex Ca_V 1.2α₁ pore-forming and auxiliary Ca_V β subunits and analyzed their trafficking dynamics from single-particle-tracking trajectories. Speed histograms obtained for each subunit were best fitted to a sum of diffusive and directed motion terms. The same mean speed for the highest-mobility state underlying directed motion was found for all subunits. The frequency of this component increased by covalent linkage of Ca_V β to Ca_V 1.2α₁ suggesting that high-speed transport occurs in association with Ca_V β. Selective tracking of Ca_V 1.2α₁ along the postendocytic pathway failed to show the highly mobile state, implying Ca_V β-independent retrograde transport. Retrograde speeds of Ca_V 1.2α₁ are compatible with myosin VI-mediated backward transport. Moreover, residency time at the cell surface was significantly prolonged when Ca_V 1.2α₁ was covalently linked to Ca_V β. Thus, Ca_V β promotes fast transport speed along anterograde trafficking and acts as a molecular switch controlling the endocytic turnover of L-type calcium channels.