title: Regulatory domain of troponin moves dynamically during activation of cardiac muscle.
abstract: Heart muscle is activated by Ca(2+) to generate force and shortening, and the signaling pathway involves allosteric mechanisms in the thin filament. Knowledge about the structure-function relationship among proteins in the thin filament is critical in understanding the physiology and pathology of the cardiac function, but remains obscure. We investigate the conformation of the cardiac troponin (Tn) on the thin filament and its response to Ca(2+) activation and propose a molecular mechanism for the regulation of cardiac muscle contraction by Tn based uniquely on information from in situ protein domain orientation. Polarized fluorescence from bifunctional rhodamine is used to determine the orientation of the major component of Tn core domain on the thin filaments of cardiac muscle. We show that the C-terminal lobe of TnC (CTnC) does not move during activation, suggesting that CTnC, together with the coiled coil formed by the TnI and TnT chains (IT arm), acts as a scaffold that holds N-terminal lobe of TnC (NTnC) and the actin binding regions of troponin I. The NTnC, on the other hand, exhibits multiple orientations during both diastole and systole. By combining the in situ orientation data with published in vitro measurements of intermolecular distances, we construct a model for the in situ structure of the thin filament. The conformational dynamics of NTnC plays an important role in the regulation of cardiac muscle contraction by moving the C-terminal region of TnI from its actin-binding inhibitory location and enhancing the movement of tropomyosin away from its inhibitory position.
authors.affiliation: Randall Division of Cell and Molecular Biophysics and British Heart Foundation Centre of Research Excellence, King's College London, London, SE1 1UL, UK.
authors.forename: Ivanka
authors.initials: I
authors.lastname: Sevrieva
authors.affiliation: Randall Division of Cell and Molecular Biophysics and British Heart Foundation Centre of Research Excellence, King's College London, London, SE1 1UL, UK.
authors.forename: Andrea C
authors.initials: AC
authors.lastname: Knowles
authors.affiliation: Randall Division of Cell and Molecular Biophysics and British Heart Foundation Centre of Research Excellence, King's College London, London, SE1 1UL, UK.
authors.forename: Thomas
authors.initials: T
authors.lastname: Kampourakis
authors.affiliation: Randall Division of Cell and Molecular Biophysics and British Heart Foundation Centre of Research Excellence, King's College London, London, SE1 1UL, UK. Electronic address: yin-biao.sun@kcl.ac.uk.
authors.forename: Yin-Biao
authors.initials: YB
authors.lastname: Sun
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000378:metabolism; D006801:Humans; D009200:Myocardial Contraction; D009206:Myocardium / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism; D017434:Protein Structure, Tertiary; D051381:Rats; D019209:Troponin C / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D019209:Troponin C; D002118:Calcium
keywords: Cardiac muscle; Contractile protein structure; Polarized fluorescence; Thin filament; Troponin
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