L-type calcium channels (LTCCs) play important roles in regulating cardiomyocyte physiology, which is governed by appropriate LTCC trafficking to and density at the cell surface. Factors influencing the expression, half-life, subcellular trafficking, and gating of LTCCs are therefore critically involved in conditions of cardiac physiology and disease.

Yeast 2-hybrid screenings, biochemical and molecular evaluations, protein interaction assays, fluorescence microscopy, structural molecular modeling, and functional studies were used to investigate the molecular mechanisms through which the LTCC Ca_vβ2 chaperone regulates channel density at the plasma membrane.

On the basis of our previous results, we found a direct linear correlation between the total amount of the LTCC pore-forming Ca_vα1.2 and the Akt-dependent phosphorylation status of Ca_vβ2 both in a mouse model of diabetic cardiac disease and in 6 diabetic and 7 nondiabetic cardiomyopathy patients with aortic stenosis undergoing aortic valve replacement. Mechanistically, we demonstrate that a conformational change in Ca_vβ2 triggered by Akt phosphorylation increases LTCC density at the cardiac plasma membrane, and thus the inward calcium current, through a complex pathway involving reduction of Ca_vα1.2 retrograde trafficking and protein degradation through the prevention of dynamin-mediated LTCC endocytosis; promotion of Ca_vα1.2 anterograde trafficking by blocking Kir/Gem-dependent sequestration of Ca_vβ2, thus facilitating the chaperoning of Ca_vα1.2; and promotion of Ca_vα1.2 transcription by the prevention of Kir/Gem-mediated shuttling of Ca_vβ2 to the nucleus, where it limits the transcription of Ca_vα1.2 through recruitment of the heterochromatin protein 1γ epigenetic repressor to the Cacna1c promoter. On the basis of this mechanism, we developed a novel mimetic peptide that, through targeting of Ca_vβ2, corrects LTCC life-cycle alterations, facilitating the proper function of cardiac cells. Delivery of mimetic peptide into a mouse model of diabetic cardiac disease associated with LTCC abnormalities restored impaired calcium balance and recovered cardiac function.

We have uncovered novel mechanisms modulating LTCC trafficking and life cycle and provide proof of concept for the use of Ca_vβ2 mimetic peptide as a novel therapeutic tool for the improvement of cardiac conditions correlated with alterations in LTCC levels and function.