The beta1a subunit regulates the functional properties of adult frog and mouse L-type Ca2+ channels of skeletal muscle.
The beta(1a) subunit, one of the auxiliary subunits of Ca(V)1.1 channels, was expressed in COS-1 cells, purified by electroelution and electrodialysis techniques and identified by Western blot using monoclonal antibodies. The purified beta(1a) subunit strongly interacted in vitro with the alpha interaction domain (AID) of Ca(V)1.1 channels. The actions of the purified beta(1a) subunit on Ca(V)1.1 channel currents were assessed in whole cell voltage clamp experiments performed in vesicles derived from frog and mouse adult skeletal muscle plasma membranes. L-type inward currents were recorded in solutions containing Ba(2+) (I(Ba)). Values of peak I(Ba) were doubled by the beta(1a) subunit in frog and mouse muscle vesicles and the amplitude of the slow component of tail currents was greatly increased. The actions of the beta(1a) subunit on Ca(V)1.1 channel currents reached a steady state within 20 min. The beta(1a) subunit had no effect on the time courses of activation or inactivation of I(Ba) or shifted the current-voltage relation. Non-linear capacitive currents were recorded in solutions that contained mostly impermeant ions. Charge movement depended on voltage with average Boltzmann parameters: Q(max) = 28.0 +/- 6.6 nC microF(-1), V = -58.0 +/- 2.0 mV and k = 15.3 +/- 1.1 mV (n = 24). In the presence of the beta(1a) subunit, these parameters remained unchanged: Q(max) = 29.8 +/- 3.5 nC microF(-1), V = -54.5 +/- 2.2 mV and k = 16.4 +/- 1.3 mV (n = 21). Overall, the work describes a novel preparation to explore in situ the role of the beta(1a) subunit on the function of adult Ca(V)1.1 channels.
García
Rubén
R
Department of Pharmacology, Centro de Investigación y de Estudios Avanzados del IPN, México DF 07360, México.
Carrillo
Elba
E
Rebolledo
Santiago
S
García
María C
MC
Sánchez
Jorge A
JA
D000465:Algorithms; D000818:Animals; D001001:Anura; D001464:Barium / Q000378:metabolism; D019556:COS Cells; D015220:Calcium Channels; D020746:Calcium Channels, L-Type / Q000235:genetics / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology; D002522:Chlorocebus aethiops; D018076:DNA, Complementary; D004058:Diffusion; D004591:Electrophoresis, Polyacrylamide Gel; D004594:Electrophysiology; D066298:In Vitro Techniques; D007700:Kinetics; D008564:Membrane Potentials / Q000502:physiology; D051379:Mice; D018482:Muscle, Skeletal / Q000502:physiology*; D018408:Patch-Clamp Techniques; D021122:Protein Subunits / Q000235:genetics / Q000502:physiology*; D014162:Transfection
C492923:Cacnb1 protein, mouse; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D018076:DNA, Complementary; D021122:Protein Subunits; D001464:Barium