The troponin C G159D mutation blunts myofilament desensitization induced by troponin I Ser23/24 phosphorylation.
Striated muscle contraction is regulated by the binding of Ca(2+) to the N-terminal regulatory lobe of the cardiac troponin C (cTnC) subunit in the troponin complex. In the heart, beta-adrenergic stimulation induces protein kinase A phosphorylation of cardiac troponin I (cTnI) at Ser23/24 to alter the interaction of cTnI with cTnC in the troponin complex and is critical to the regulation of cardiac contractility. We investigated the effect of the dilated cardiomyopathy linked cTnC Gly159 to Asp (cTnC-G159D) mutation on the development of Ca(2+)-dependent tension and ATPase rate in whole troponin-exchanged skinned rat trabeculae. Even though this mutation is located in the C-terminal lobe of cTnC, the G159D mutation was demonstrated to depress ATPase activation and filament sliding in vitro. The effects of this mutation within the cardiac myofilament are unknown. Our results demonstrate that the cTnC-G159D mutation by itself does not alter the myofilament response to Ca(2+) in the cardiac muscle lattice. However, in the presence of cTnI phosphorylated at Ser23/24, which reduced Ca(2+) sensitivity and enhanced cross-bridge cycling in controls, cTnC-G159D specifically blunted the phosphorylation induced decrease in Ca(2+)-sensitive tension development without altering cross-bridge cycling. Measurements in purified troponin confirmed that this cTnC-G159D blunting of myofilament desensitization results from altered Ca(2+)-binding to cTnC. Our results provide novel evidence that modification of the cTnC-cTnI interaction has distinct effects on troponin Ca(2+)-binding and cross-bridge kinetics to suggest a novel role for thin filament mutations in the modulation of myofilament function through beta-adrenergic signaling as well as the development of cardiomyopathy.
Biesiadecki
Brandon J
BJ
Department of Physiology and Biophysics and Center for Cardiovascular Research, College of Medicine, University of Illinois, Chicago, IL 60612, USA.
Kobayashi
Tomoyoshi
T
Walker
John S
JS
Solaro
R John
RJ
de Tombe
Pieter P
PP
D008841:Actin Cytoskeleton / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D009202:Cardiomyopathies / Q000209:etiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology; D006801:Humans; D009154:Mutation; D010766:Phosphorylation; D011943:Receptors, Adrenergic, beta / Q000502:physiology; D012694:Serine; D019209:Troponin C / Q000235:genetics*; D019210:Troponin I / Q000378:metabolism*
D011943:Receptors, Adrenergic, beta; D019209:Troponin C; D019210:Troponin I; D012694:Serine; D017868:Cyclic AMP-Dependent Protein Kinases; D002118:Calcium