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Dissection of functional domains of the voltage-dependent Ca2+ channel alpha2delta subunit.
Felix R
,
Gurnett CA
,
De Waard M
,
Campbell KP
.
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Coexpression of the cloned voltage-dependent Ca2+ channel alpha2delta subunit with the pore-forming alpha1 subunit results in a significant increase in macroscopic current amplitude. To gain insight into the mechanism underlying this interaction, we have examined the regulatory effect of either the alpha2delta complex or the delta subunit on the Ca2+ channel alpha1 subunit. Transient transfection of tsA201 cells with the cardiac L-type alpha1C subunit alone resulted in the expression of inward voltage-activated currents as well as measurable [3H]-PN200-110 binding to membranes from transfected cells. Coexpression of the alpha2delta subunit significantly increased the macroscopic current amplitude, altered the voltage dependence and the kinetics of the current, and enhanced [3H]-PN200-110 binding. Except for the increase in amplitude, coexpression of the delta subunit reproduced entirely the effects of the full-length alpha2delta subunit on the biophysical properties of the alpha1C currents. However, no effect on specific [3H]-PN200-110 binding was observed on delta subunit coexpression. Likewise, profound effects on current kinetics of the neuronal alpha1A subunit were observed on coexpression of the alpha2delta complex in Xenopus oocytes. Furthermore, by using a chimeric strategy, we localized the region involved in this regulation to the transmembrane domain of the delta subunit. These data strongly suggest that the molecular determinants involved in alpha2delta regulation are conserved across L-type and non-L type Ca2+ channels. Taken together, our results indicate that the region of the alpha2delta subunit involved in the modulation of the gating properties of the high voltage-activated calcium channels is localized in the delta domain of the protein. In contrast, the level of membrane expression of functional channels relies on the presence of the alpha2 domain of the alpha2delta complex.
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