Modification of Ca2+ channel activity by deletions at the carboxyl terminus of the cardiac alpha 1 subunit.
Voltage-sensitive Ca2+ channels are multisubunit complexes that include, among others, a large alpha 1 subunit, which by itself is sufficient to form a channel. Several alpha 1 genes encoding L-, N-, and P-type Ca2+ channels have been cloned. These alpha 1 genes share a high degree of sequence homology in the putative transmembrane regions, but vary substantially in the putative intracellular loops and the flanking amino and carboxyl termini. In the present study, we investigated the functional roles of the 665-amino acid long carboxyl terminus of a cardiac alpha 1 by constructing deletion mutants. Expression in Xenopus oocytes of delta C1856, delta C1733, and delta C1700, which lack from 307 to 472 amino acids at the carboxyl terminus, led to inward Ba2+ currents that were 4- to 6-fold greater than observed with the 2171-amino acid long wild type alpha 1. Ionic currents increased without a change in the amount of charge moved during voltage-dependent gating, suggesting that the increase in ionic currents was not due to an increase in the number of channels that were expressed. Single channel analysis revealed an unaltered unitary conductance. Thus, removal of up to 70% of the carboxyl terminus increased current density by facilitating the coupling between the voltage-dependent gating and channel opening, leading to an increased opening probability of the channel.
Wei
X
X
Department of Molecular Physiology, Baylor College of Medicine, Houston, Texas 77030.
Neely
A
A
Lacerda
A E
AE
Olcese
R
R
Stefani
E
E
Perez-Reyes
E
E
Birnbaumer
L
L
D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester / Q000494:pharmacology; D000818:Animals; D001464:Barium / Q000378:metabolism; D001483:Base Sequence; D015220:Calcium Channels / Q000096:biosynthesis / Q000187:drug effects / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers; D017275:Isradipine / Q000378:metabolism; D046911:Macromolecular Substances; D008564:Membrane Potentials / Q000187:drug effects; D008969:Molecular Sequence Data; D009206:Myocardium / Q000378:metabolism*; D016376:Oligonucleotides, Antisense; D009865:Oocytes / Q000187:drug effects / Q000502:physiology; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000187:drug effects / Q000378:metabolism; D015183:Restriction Mapping; D017384:Sequence Deletion; D014162:Transfection; D014982:Xenopus laevis
D015220:Calcium Channels; D017931:DNA Primers; D046911:Macromolecular Substances; D016376:Oligonucleotides, Antisense; D011994:Recombinant Proteins; D001464:Barium; D001498:3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethyl-5-nitro-4-(2-(trifluoromethyl)phenyl)-, Methyl ester; D017275:Isradipine