Structural features determining differential receptor regulation of neuronal Ca channels

Dihydropyridine-insensitive Ca channels are subject to direct receptor G-protein-mediated inhibition to differing extents. alpha1B channels are much more strongly modulated than alpha1E channels. To understand the structural basis for this difference, we have constructed and expressed various alpha1B and alpha1E chimeric Ca channels and examined their regulation by kappa-opioid receptors. Replacement of the first membrane-spanning domain of alpha1E with the corresponding region of alpha1B resulted in a chimeric Ca channel that was modulated by kappa-opioid receptors to a significantly greater extent than alpha1E. Transfer of the N terminus and I/II loop from alpha1B in addition to domain I resulted in a chimeric channel that was modulated to the same extent as alpha1B. Other regions of the molecule do not appear to contribute significantly to the degree of inhibition obtained, although the C terminus may contribute to facilitation.