State- and isoform-dependent interaction of isradipine with the alpha1C L-type calcium channel.
We investigated the dihydropyridine (DHP) inhibition of barium current (I(Ba)) through the smooth muscle alpha1Ch and cardiac alpha1Ca splice variants of the L-type calcium channel using a whole-cell patch-clamp method. IC50 values for inhibition of current amplitude of the alpha1Cb channel were three to fivefold lower than for the alpha1Ca channel at holding potentials between -80 mV and -30 mV. No difference was found in either the transition of the channels into an inactivated state in the absence or presence of drug, or in the recovery from inactivation under control conditions. However, isradipine slowed the recovery from inactivation of alpha1Ca more effectively than alpha1Cb. To evaluate the interaction of isradipine with the open channel state of both splice variants, interactions with the inactivated state were selectively suppressed by the mutation of three amino acids in the IVS6 segment (Y1485I, M1486F, I1493L) in alpha1CbCh30 and alpha1CaCh30 channels. The extent of this interaction was seen by an acceleration of current decay. This was found to be identical for both splice variants. Our results suggest that the higher DHP selectivity of the alpha1Cb versus the alpha1Ca channel is caused by the structural difference in the binding site and not by different transitions between resting, open and inactivated states.
Lacinová
L
L
Institut für Pharmakologie und Toxikologie der Technischen Universität München, Munich, Germany. lacinova@ipt.med.tu-muenchen.de
Klugbauer
N
N
Hofmann
F
F
D017398:Alternative Splicing / Q000235:genetics; D001464:Barium / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000187:drug effects* / Q000378:metabolism*; D002460:Cell Line; D004095:Dihydropyridines / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D017136:Ion Transport / Q000187:drug effects / Q000235:genetics; D017275:Isradipine / Q000494:pharmacology*; D008564:Membrane Potentials / Q000187:drug effects; D009130:Muscle, Smooth / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009206:Myocardium / Q000378:metabolism; D018408:Patch-Clamp Techniques; D020033:Protein Isoforms / Q000187:drug effects / Q000378:metabolism; D014162:Transfection
D020746:Calcium Channels, L-Type; D004095:Dihydropyridines; D020033:Protein Isoforms; D001464:Barium; C038806:1,4-dihydropyridine; D017275:Isradipine