title: Protein kinase A mediated modulation of acto-myosin kinetics.
abstract: The effects of protein kinase A (PKA) mediated phosphorylation on thin filament and cross-bridge function is not fully understood. To delineate the effects of troponin I (TnI) phosphorylation by PKA on contractile protein performance, reconstituted thin filaments were treated with PKA. With the use of the in vitro motility assay, PKA treated thin filament function was assessed relative to non-phosphorylated thin filaments in a calcium-regulated system. At maximal calcium activation, unloaded shortening velocity and force did not differ between the groups. However, at submaximal activation, an increase in calcium sensitivity of the thin filament was observed for velocity but a decrease in calcium sensitivity was observed for force. Activation of the thin filament by myosin strong-binding did not elicit a calcium-independent effect. The rightward shift in calcium sensitivity for force and the leftward shift in calcium sensitivity for velocity indicate that PKA phosphorylation of TnI directly modulates the kinetics of the myosin cross-bridge. In addition, the altered velocity dependence on thin filament length implicates reduced myosin cross-bridge binding with PKA treatment. These data highlight the importance of TnI serine 23 and 24 phosphorylation in the modulation of cardiac function.
authors.affiliation: Cardiology Unit, Department of Medicine, College of Medicine, University of Vermont, 126C HSRF Building, 149 Beaumont Avenue, Burlington, VT 05405, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Hünlich
authors.forename: Kelly J
authors.initials: KJ
authors.lastname: Begin
authors.forename: Joseph A
authors.initials: JA
authors.lastname: Gorga
authors.forename: David E
authors.initials: DE
authors.lastname: Fishbaugher
authors.forename: Martin M
authors.initials: MM
authors.lastname: LeWinter
authors.forename: Peter
authors.initials: P
authors.lastname: VanBuren
mesh_terms: D000205:Actomyosin / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism / Q000494:pharmacology; D002645:Chickens; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D007700:Kinetics; D010766:Phosphorylation; D019210:Troponin I / Q000378:metabolism
chemical_list: D019210:Troponin I; D000205:Actomyosin; D017868:Cyclic AMP-Dependent Protein Kinases; D002118:Calcium