The role of dynamic palmitoylation in Ca2+ channel inactivation
Abstract
N- and P/Q-type Ca2+ channels regulate a number of critical physiological processes including synaptic transmission and hormone secretion. These Ca2+ channels are multisubunit proteins, consisting of a pore-forming α1, and accessory β and α2δ subunits each encoded by multiple genes and splice variants. β subunits alter current amplitude and kinetics. The β2a subunit is associated with slowed inactivation, an effect that requires the palmitoylation of two N-terminal cysteine residues in β2a. In the current manuscript, we studied steady state inactivation properties of native N- and P/Q-type Ca2+ channels and recombinant N-type Ca2+ channels. When bovine α1B and β2a and human α2δ were coexpressed in tsA 201 cells, we observed significant variations in inactivation; some cells exhibited virtually no inactivation as the holding potential was altered whereas others exhibited significant inactivation. A similar variability in inactivation was observed in native channels from bovine chromaffin cells. In individual chromaffin cells, the amount of inactivation exhibited by N-type channels was correlated with the inactivation of P/Q-type channels, suggesting a shared mechanism. Our results with recombinant channels with known β subunit composition indicated that inactivation could be dynamically regulated, possibly by alterations in β subunit palmitoylation. Tunicamycin, which inhibits palmitoylation, increased steady-state inactivation of Ca2+ channels in chromaffin cells. Cerulenin, another drug that inhibits palmitoylation, also increased inactivation. Tunicamycin produced a similar effect on recombinant N-type Ca2+ channels containing β2a but not β2b or β2a subunits mutated to be palmitoylation deficient. Our results suggest that Ca2+ channels containing β2a subunits may be regulated by dynamic palmitoylation.
- Publication:
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Proceedings of the National Academy of Science
- Pub Date:
- August 2000
- DOI:
- 10.1073/pnas.160589697
- Bibcode:
- 2000PNAS...97.9293H
- Keywords:
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- Neurobiology