The interplay between splicing of exons 18a and 47 differentially affects membrane targeting and function of human Ca _V 2.2

N-type calcium channels (Ca _V 2.2) are predominantly localized in presynaptic terminals, and are particularly important for pain transmission in the spinal cord. Furthermore, they have multiple isoforms, conferred by alternatively-spliced or cassette exons, which are differentially expressed. Here we have examined alternatively-spliced exon47 variants that encode a long or short C-terminus in human Ca _V 2.2. In the Ensembl database, all short exon47-containing transcripts were associated with the absence of exon 18a, therefore we also examined effect of inclusion or absence of exon18a, combinatorially with the exon47 splice variants. We found that long exon47, only in the additional presence of exon18a, results in Ca _V 2.2 currents that have a 3.6-fold greater maximum conductance than the other three combinations. In contrast, cell surface expression of Ca _V 2.2 in both tsA-201 cells and hippocampal neurons is increased ∼4-fold by long exon47 relative to short exon47, in either the presence or absence of exon18a. This surprising discrepancy between trafficking and function indicates that cell surface expression is enhanced by long exon47, independently of exon 18a. However, in the presence of exon47, exon18a mediates an additional permissive effect on Ca _V 2.2 gating. We also investigated the SNP in exon47 that has been linked to schizophrenia and Parkinson’s disease, which we found is only non-synonymous in the short exon47 C-terminal isoform, resulting in two minor alleles. This study highlights the importance of investigating the combinatorial effects of exon inclusion, rather than each in isolation, in order to increase our understanding of calcium channel function.