Alpha-adrenergic receptor stimulation in rat ventricular myocytes phosphorylates MYPT via Rho kinase, dissociating it from PP1 to dynamically modulate LC2 phosphorylation.
Does alpha-adrenergic receptor stimulation modulate MYPT and PP1 to regulate LC2 phosphorylation in adult rat ventricular myocytes?
Alpha-adrenergic receptor activation works through Rho kinase to phosphorylate MYPT, leading to its dissociation from PP1 and dynamic modulation of LC2 phosphorylation, which regulates myocardial function.
Myosin light chain 2 (LC2) phosphorylation is of both physiological and pathological importance to myocardial function. The phosphatase that directly dephosphorylates LC2 is a type 1 protein phosphatase (PP1) that contains a catalytic subunit that complexes with a myosin-binding phosphatase targeting subunit (MYPT). The goal of the present study was to examine the role of MYPT in the regulation of PP1 in ventricular myocytes. In the first part of the study, regional distribution of MYPT expression and phosphorylation were determined in unstimulated hearts. The pattern of MYPT phosphorylation was inversely related to the LC2 phosphorylation spatial gradient as described by Epstein and colleagues (Davis JS, Hassanzadeh S, Winitsky S, Lin H, Satorius C, Vemuri R, Aletras AH, Wen H, and Epstein ND. Cell 107: 631-641, 2001). In the second part of the study, adult rat isolated ventricular myocytes were exposed to an alpha-adrenergic receptor agonist, and properties of MYPT, PP1, and LC2 were studied. We found MYPT associates with cardiac myofilaments, and this association increases upon alpha-adrenergic receptor stimulation. Activation of alpha-adrenergic receptors also led to a decrease in the PP1-myofilament association. Furthermore, alpha-adrenergic receptor stimulation results in phosphorylation of MYPT and LC2 and an increase in myocyte Ca(2+) sensitivity of tension that all depend on Rho kinase activation. These data support the hypothesis that alpha-adrenergic receptor activation works through Rho kinase to phosphorylate MYPT, and phosphorylated MYPT dissociates from PP1 so that PP1 is no longer physically associated with LC2. Hence, we propose a pathway for the dynamic modulation of LC2 phosphorylation through receptor-dependent phosphorylation of MYPT, and a spatial gradient of LC2 phosphorylation under basal conditions that occurs due to varied levels of phosphorylation of MYPT in ventricles.
Ravi et al. (2005) studied Myocardial function. Alpha-adrenergic receptor agonist vs. Unstimulated conditions was evaluated on MYPT expression, phosphorylation, and association with PP1 and LC2. Alpha-adrenergic receptor stimulation in rat ventricular myocytes phosphorylates MYPT via Rho kinase, dissociating it from PP1 to dynamically modulate LC2 phosphorylation.