Isoprenaline stimulation phosphorylated TnI, C-protein, and phospholamban in rabbit hearts, with phospholamban playing a dominant role in the mechanical response to beta-adrenergic stimulation.
Demonstrates that phospholamban phosphorylation is the primary driver of the mechanical response to beta-adrenergic stimulation in the mammalian heart, rather than TnI or C-protein phosphorylation.
Phosphorylation of myofibrillar and sacroplasmic-reticulum (SR) proteins was studied in Langendorff-perfused rabbit hearts subjected to various inotropic interventions. Stimulation of hearts with isoprenaline resulted in the phosphorylation of both troponin I (TnI) and C-protein in myofibrils and phospholamban in SR. Phosphorylation of phospholamban could be reversed by a 15 min perfusion with drug-free buffer, after a 1 minute pulse perfusion with isoprenaline, at which time the mechanical effects of isoprenaline stimulation had also been reversed. However, both TnI and C-protein remained phosphorylated at this time. Moreover, the inhibition of Ca2+ activation of the Mg2+-dependent ATPase (Mg-ATPase) activity associated with myofibrillar phosphorylation persisted in myofibrils prepared from hearts frozen after 15 min of washout of isoprenaline. To assess the contribution of C-protein phosphorylation in the decrease of Ca2+ activation of the myofibrillar Mg-ATPase activity, we reconstituted a regulated actomyosin system in which only C-protein was phosphorylated. In this system, C-protein phosphorylation did not contribute to the decrease in Ca2+ activation of Mg-ATPase activity, indicating that TnI phosphorylation is responsible for the diminished sensitivity of the myofibrils to Ca2+. These observations support the hypothesis that phospholamban phosphorylation plays a more dominant role than TnI or C-protein phosphorylation in the mechanical response of the mammalian heart to beta-adrenergic stimulation.
Garvey et al. (1988) studied Isolated rabbit hearts. Isoprenaline vs. Drug-free buffer was evaluated on Phosphorylation of myofibrillar and sacroplasmic-reticulum (SR) proteins and Ca2+ activation of Mg-ATPase activity. Isoprenaline stimulation phosphorylated TnI, C-protein, and phospholamban in rabbit hearts, with phospholamban playing a dominant role in the mechanical response to beta-adrenergic stimulation.