title: Post-translational modifications of myofilament proteins involved in length-dependent prolongation of relaxation in rabbit right ventricular myocardium.
abstract: The phosphorylation state of several cardiac myofilament proteins changes with the level of stretch in intact, twitch-contracting cardiac muscles. It remains unclear which kinases are involved in the length-dependent phosphorylation of these proteins. We set out to investigate which kinases are involved after a step-wise change in cardiac muscle length. We hypothesize that myofilament protein phosphorylation by PKCβII and PKA alters contractile kinetics during length-dependent activation. Right ventricular intact trabeculae were isolated from New Zealand White rabbit hearts and stimulated to contract at 1Hz. Twitch force recordings where taken at taut and optimal muscle lengths before and after administration of kinase inhibitors at 37°C. PKCβII inhibition significantly decreased time from stimulation to peak force (TTP), time from peak force to 50% relaxation (RT50), and 90% relaxation (RT90) at optimal muscle length. This led to a loss in the length-dependent increase of RT50 and RT90 in the presence of the PKCβII inhibitor, whereas the length-dependent increase in RT50 and RT90 was seen in the controls. PKA inhibition using H-89 significantly decreased TTP at both taut and optimal muscle lengths. Detection of Ser/Thr phosphorylation with ProQ-diamond staining indicates a role for PKCβII in the phosphorylation of tropomyosin and myosin light chain-2 (MLC2) and PKA for tropomyosin, troponin-I, MLC2, myosin binding protein-C, troponin-T (TnT) 3 and TnT4. Our data provide evidence for two signaling kinases acting upon myofilament proteins during length-dependent activation, and provide further insight for length-dependent myofilament function.
authors.affiliation: Department of Physiology and Cell Biology, College of Medicine and D. Davis Heart Lung Institute, The Ohio State University, 1645 Neil Avenue, Columbus, OH 43210-1218, USA.
authors.forename: Michelle M
authors.initials: MM
authors.lastname: Monasky
authors.forename: Domenico M
authors.initials: DM
authors.lastname: Taglieri
authors.forename: Alice K
authors.initials: AK
authors.lastname: Jacobson
authors.forename: Kaylan M
authors.initials: KM
authors.lastname: Haizlip
authors.forename: R John
authors.initials: RJ
authors.lastname: Solaro
authors.forename: Paul M L
authors.initials: PM
authors.lastname: Janssen
mesh_terms: D000818:Animals; D024722:Cardiac Myosins / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism; D006352:Heart Ventricles / Q000378:metabolism*; D007546:Isoquinolines / Q000494:pharmacology; D008297:Male; D009200:Myocardial Contraction; D009206:Myocardium / Q000378:metabolism*; D009210:Myofibrils / Q000378:metabolism*; D018994:Myosin Light Chains / Q000378:metabolism; D010766:Phosphorylation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D064546:Protein Kinase C beta; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011499:Protein Processing, Post-Translational; D011817:Rabbits; D013194:Staining and Labeling / Q000379:methods; D019311:Staurosporine / Q000494:pharmacology; D013449:Sulfonamides / Q000494:pharmacology; D013997:Time Factors; D014335:Tropomyosin / Q000378:metabolism; D019210:Troponin I / Q000378:metabolism; D020107:Troponin T / Q000378:metabolism
chemical_list: D007546:Isoquinolines; D018994:Myosin Light Chains; D047428:Protein Kinase Inhibitors; D013449:Sulfonamides; D014335:Tropomyosin; D019210:Troponin I; D020107:Troponin T; C107184:myosin light chain 2; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D024722:Cardiac Myosins; D019311:Staurosporine; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide