title: Dynamics of PKA phosphorylation and gain of function in cardiac pacemaker cells: a computational model analysis.
abstract: Cardiac pacemaker cell function is regulated by a coupled-clock system that integrates molecular cues on the cell-membrane surface (i.e., membrane clock) and on the sarcoplasmic reticulum (SR) (i.e., Ca(2+) clock). A recent study has shown that cotransfection of spontaneous beating cells (HEK293 cells and neonatal rat myocytes) with R524Q-mutant human hyperpolarization-activated cyclic nucleotide-gated molecules (the dominant component of funny channels) increases the funny channel's sensitivity to cAMP and leads to a decrease in spontaneous action potential (AP) cycle length (i.e., tachycardia). We hypothesize that in rabbit pacemaker cells, the same behavior is expected, and because of the coupled-clock system, the resultant steady-state decrease in AP cycle length will embody contributions from both clocks: the initial decrease in the spontaneous AP beating interval, arising from increased sensitivity of the f-channel to cAMP, will be accompanied by an increase in the adenylyl cyclase (AC)-cAMP-PKA-dependent phosphorylation activity, which will further decrease this interval. To test our hypothesis, we used the recently developed Yaniv-Lakatta pacemaker cell numerical model. This model predicts the cAMP signaling dynamics, as well as the kinetics and magnitude of protein phosphorylation in both normal and mutant pacemaker cells. We found that R524Q-mutant pacemaker cells have a shorter AP firing rate than that of wild-type cells and that gain in pacemaker function is the net effect of the R514Q mutation on the functioning of the coupled-clock system. Specifically, our results directly support the hypothesis that changes in Ca(2+)-activated AC-cAMP-PKA signaling are involved in the development of tachycardia in R524Q-mutant pacemaker cells.
authors.affiliation: Laboratory of Bioenergetic and Bioelectric Systems, Biomedical Engineering Faculty, Technion-IIT, Haifa, Israel.
authors.forename: Joachim
authors.initials: J
authors.lastname: Behar
authors.affiliation: Laboratory of Bioenergetic and Bioelectric Systems, Biomedical Engineering Faculty, Technion-IIT, Haifa, Israel yaely@bm.technion.ac.il.
authors.forename: Yael
authors.initials: Y
authors.lastname: Yaniv
mesh_terms: D000200:Action Potentials; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001683:Biological Clocks; D002135:Calcium-Binding Proteins / Q000378:metabolism; D003198:Computer Simulation; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D020022:Genetic Predisposition to Disease; D006339:Heart Rate; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008955:Models, Cardiovascular; D009154:Mutation; D009716:Numerical Analysis, Computer-Assisted; D010641:Phenotype; D010766:Phosphorylation; D011817:Rabbits; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases / Q000378:metabolism; D015290:Second Messenger Systems; D012849:Sinoatrial Node / Q000166:cytology / Q000201:enzymology*; D013610:Tachycardia / Q000201:enzymology* / Q000235:genetics / Q000503:physiopathology; D000099211:Phospholamban
chemical_list: D000262:Adenylyl Cyclases; D002135:Calcium-Binding Proteins; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D064428:Hyperpolarization-Activated Cyclic Nucleotide-Gated Channels; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000099211:Phospholamban
keywords: Ca2+ cycling; Ca2+-excitation contraction coupling; arrhythmia; pacemaker cell automaticity