Role of CaVbeta subunits, and lack of functional reserve, in protein kinase A modulation of cardiac CaV1.2 channels.
Protein kinase A (PKA)-mediated enhancement of L-type calcium currents (I(Ca,L)) is essential for sympathetic regulation of the heartbeat and is the classic example of channel regulation by phosphorylation, and its loss is a common hallmark of heart failure. Mechanistic understanding of how distinct Ca(V) channel subunits contribute to PKA modulation of I(Ca,L) has been intensely pursued yet remains elusive. Moreover, critical features of this regulation such as its functional reserve (the surplus capacity available for modulation) in the heart are unknown. Here, we use an overexpression paradigm in heart cells to simultaneously identify the impact of auxiliary Ca(V)betas on PKA modulation of I(Ca,L) and to gauge the functional reserve of this regulation in the heart. Ca(V)1.2 channels containing wild-type beta(2a) or a phosphorylation-deficient mutant (beta(2a,AAA)) were equally upregulated by PKA, discounting a necessary role for beta phosphorylation. Nevertheless, channels reconstituted with beta(2a) displayed a significantly diminished PKA response compared with other beta isoforms, an effect explainable by a uniquely higher basal P(o) of beta(2a) channels. Overexpression of all betas increased basal current density, accompanied by a concomitant decrease in the magnitude of PKA regulation. Scatter plots of fold increase in current against basal current density revealed an inverse relationship that was conserved across species and conformed to a model in which a large fraction of channels remained unmodified after PKA activation. These results redefine the role of beta subunits in PKA modulation of Ca(V)1.2 channels and uncover a new design principle of this phenomenon in the heart, vis à vis a limited functional reserve.
Miriyala
Jayalakshmi
J
Columbia University, College of Physicians and Surgeons, Department of Physiology and Cellular Biophysics, 630 W 168th St, New York, NY 10032, USA.
Nguyen
Trang
T
Yue
David T
DT
Colecraft
Henry M
HM
D000818:Animals; D002118:Calcium / Q000378:metabolism; D020746:Calcium Channels, L-Type / Q000378:metabolism*; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004594:Electrophysiology; D006168:Guinea Pigs; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D018408:Patch-Clamp Techniques; D010766:Phosphorylation; D021122:Protein Subunits / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley
D020746:Calcium Channels, L-Type; C419385:L-type calcium channel alpha(1C); D021122:Protein Subunits; C493217:calcium channel, l type beta2c subunit, rat; D017868:Cyclic AMP-Dependent Protein Kinases; D002118:Calcium