The α2δ Ligand Gabapentin Inhibits the Rab11-Dependent Recycling of the Calcium Channel Subunit α2δ-2

Abstract

The α2δ subunits of voltage-gated calcium channels are important modulatory subunits that enhance calcium currents and may also have other roles in synaptogenesis. The antiepileptic and antiallodynic drug gabapentin (GBP) binds to the α2δ-1 and α2δ-2 isoforms of this protein, and its binding may disrupt the binding of an endogenous ligand, required for their correct function. We have shown previously that GBP produces a chronic inhibitory effect on calcium currents by causing a reduction in the total number of α2δ and α1 subunits at the cell surface. This action of GBP is likely to be attributable to a disruption of the trafficking of α2δ subunits, either to or from the plasma membrane. We studied the effect of GBP on the internalization of, and insertion into the plasma membrane of α2δ-2 using an α-bungarotoxin binding site-tagged α2δ-2 subunit, and a fluorescent derivative of α-bungarotoxin. We found that GBP specifically disrupts the insertion of α2δ-2 from post-Golgi compartments to the plasma membrane, and this effect was prevented by a mutation of α2δ-2 that abolishes its binding to GBP. The coexpression of the GDP-bound Rab11 S25N mutant prevented the GBP-induced decrease in α2δ-2 cell surface levels, both in cell lines and in primary neurons, and the GBP-induced reduction in calcium channel currents. In contrast, the internalization of α2δ-2 was unaffected by GBP. We conclude that GBP acts by preventing the recycling of α2δ-2 from Rab11-positive recycling endosomes to the plasma membrane.