Snapin Specifically Up-Regulates Ca_v1.3 Ca²⁺ Channel Variant with a Long Carboxyl Terminus

Ca²⁺ entry through Ca_v1.3 Ca²⁺ channels plays essential roles in diverse physiological events. We employed yeast-two-hybrid (Y2H) assays to mine novel proteins interacting with Ca_v1.3 and found Snapin2, a synaptic protein, as a partner interacting with the long carboxyl terminus (CT_L) of rat Ca_v1.3_L variant. Co-expression of Snapin with Ca_v1.3_L/Ca_vβ₃/α₂δ₂ subunits increased the peak current density or amplitude by about 2-fold in HEK-293 cells and Xenopus oocytes, without affecting voltage-dependent gating properties and calcium-dependent inactivation. However, the Snapin up-regulation effect was not found for rat Ca_v1.3_S containing a short CT (CT_S) in which a Snapin interaction site in the CT_L was deficient. Luminometry and electrophysiology studies uncovered that Snapin co-expression did not alter the membrane expression of HA tagged Ca_v1.3_L but increased the slope of tail current amplitudes plotted against ON-gating currents, indicating that Snapin increases the opening probability of Ca_v1.3_L. Taken together, our results strongly suggest that Snapin directly interacts with the CT_L of Ca_v1.3_L, leading to up-regulation of Ca_v1.3_L channel activity via facilitating channel opening probability.