C-terminal fragments of the α1C (Cav1.2) subunit associate with and regulate L-type calcium channels containing C-terminal-truncated α1C subunits

L-type Ca2+ channels in native tissues have been found to contain a pore-forming alpha (1) subunit that is often truncated at the C terminus. However, the C terminus contains many important domains that regulate channel function. To test the hypothesis that C-terminal fragments may associate with and regulate C-terminal-truncated alpha (1C) (Ca(V)1.2) subunits, we performed electrophysiological and biochemical experiment, In tsA201 cells expressing either wild type or C-terminal-truncated alpha (1C) subunits in combination with a beta (2a) subunit, truncation of the alpha (1C) subunit by as little as 147 amino acids led to a 10-15-fold increase in currents compared with those obtained from control, full-length alpha (1C) subunits, Dialysis of cells expressing the truncated alpha (1C) subunits with C-terminal fragments applied through the patch pipette reconstituted the inhibition of the channels seen with full-length alpha (1C) Subunits. In addition, C-terminal deletion mutants containing a tethered C terminus also exhibited the C-terminal-induced inhibition. Immunoprecipitation assays demonstrated the association of the C-terminal fragments with truncated alpha (1C) subunits, In addition, glutathione S-transferase pull-down assays demonstrated that the C-terminal inhibitory fragment could associate with at least two domains within the C terminus. The results support the hypothesis the C-terminal fragments of the alpha (1C) subunit can associate with C-terminal-truncated alpha (1C) subunits and inhibit the currents through L-type Ca2+ channels.