p38-MAPK induced dephosphorylation of alpha-tropomyosin is associated with depression of myocardial sarcomeric tension and ATPase activity.
Our objective in work presented here was to understand the mechanisms by which activated p38alpha MAPK depresses myocardial contractility. To test the hypothesis that activation of p38 MAPK directly influences sarcomeric function, we used transgenic mouse models with hearts in which p38 MAPK was constitutively turned on by an upstream activator (MKK6bE). These hearts demonstrated a significant depression in ejection fraction after induction of the transgene. We also studied hearts of mice expressing a dominant negative p38alpha MAPK. Simultaneous determination of tension and ATPase activity of detergent-skinned fiber bundles from left ventricular papillary muscle demonstrated a significant inhibition of both maximum tension and ATPase activity in the transgenic-MKK6bE hearts. Fibers from hearts expressing dominant negative p38alpha MAPK demonstrated no significant change in tension or ATPase activity. There were no significant changes in phosphorylation level of troponin-T3 and troponin-T4, or myosin light chain 2. However, compared with controls, there was a significant depression in levels of phosphorylation of alpha-tropomyosin and troponin I in fiber bundles from transgenic-MKK6bE hearts, but not from dominant negative p38alpha MAPK hearts. Our experiments also showed that p38alpha MAPK colocalizes with alpha-actinin at the Z-disc and complexes with protein phosphatases (PP2alpha, PP2beta). These data are the first to indicate that chronic activation of p38alpha MAPK directly depresses sarcomeric function in association with decreased phosphorylation of alpha-tropomyosin.
Vahebi
Susan
S
Department of Physiology and Biophysics, Center for Cardiovascular Research, College of Medicine, University of Illinois at Chicago, 835 S. Wolcott Ave., Chicago, IL 60612-7342, USA.
Ota
Asuka
A
Li
Manxiang
M
Warren
Chad M
CM
de Tombe
Pieter P
PP
Wang
Yibin
Y
Solaro
R John
RJ
D008841:Actin Cytoskeleton / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D020013:Calcium Signaling; D064211:Connectin; D004789:Enzyme Activation; D004790:Enzyme Induction / Q000235:genetics; D055551:HSP27 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000032:analysis; D047468:Immunoprecipitation; D007537:Isometric Contraction / Q000502:physiology; D048669:MAP Kinase Kinase 6 / Q000235:genetics / Q000502:physiology; D013058:Mass Spectrometry; D051379:Mice; D008822:Mice, Transgenic; D008955:Models, Cardiovascular; D018832:Molecular Chaperones; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D009124:Muscle Proteins / Q000302:isolation & purification / Q000378:metabolism; D016297:Mutagenesis, Site-Directed; D009200:Myocardial Contraction / Q000502:physiology*; D009218:Myosins / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000032:analysis; D010210:Papillary Muscles / Q000502:physiology; D010749:Phosphoprotein Phosphatases / Q000302:isolation & purification / Q000378:metabolism; D010766:Phosphorylation; D025941:Protein Interaction Mapping; D000071636:Protein Phosphatase 2C; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000502:physiology; D012518:Sarcomeres / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D019076:Transgenes; D014335:Tropomyosin / Q000378:metabolism*; D019210:Troponin I / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*
D064211:Connectin; D055551:HSP27 Heat-Shock Proteins; D006360:Heat-Shock Proteins; C486113:Hsbp1 protein, mouse; C486699:Hspb2 protein, mouse; D018832:Molecular Chaperones; D009097:Multienzyme Complexes; D009124:Muscle Proteins; D009363:Neoplasm Proteins; D011993:Recombinant Fusion Proteins; D014335:Tropomyosin; D019210:Troponin I; D048051:p38 Mitogen-Activated Protein Kinases; D048669:MAP Kinase Kinase 6; D010749:Phosphoprotein Phosphatases; D000071636:Protein Phosphatase 2C; D009218:Myosins; D002118:Calcium