Age-Related Homeostatic Midchannel Proteolysis of Neuronal L-type Voltage-Gated Ca²⁺ Channels

Highlights

• The core of L-type VGCC pore-forming subunit Ca_v1.2 undergoes regulated cleavage

• Cleaved Ca_v1.2 fragments stay on the plasma membrane but can dissociate

• Aging, channel activity, calpain, and PEST sequences play a role in Ca_v1.2 proteolysis

• Ca_v1.2 core cleavage greatly changes L-type VGCC properties and currents

Summary

Neural circuitry and brain activity depend critically on proper function of voltage-gated calcium channels (VGCCs), whose activity must be tightly controlled. We show that the main body of the pore-forming α₁ subunit of neuronal L-type VGCCs, Ca_v1.2, is proteolytically cleaved, resulting in Ca_v1.2 fragment channels that separate but remain on the plasma membrane. This “midchannel” proteolysis is regulated by channel activity, involves the Ca²⁺-dependent protease calpain and the ubiquitin-proteasome system, and causes attenuation and biophysical alterations of VGCC currents. Recombinant Ca_v1.2 fragment channels mimicking the products of midchannel proteolysis do not form active channels on their own but, when properly paired, produce currents with distinct biophysical properties. Midchannel proteolysis increases dramatically with age and can be attenuated with an L-type VGCC blocker in vivo. Midchannel proteolysis represents a novel form of homeostatic negative-feedback processing of VGCCs that could profoundly affect neuronal excitability, neurotransmission, neuroprotection, and calcium signaling in physiological and disease states.