Direct interaction between troponin and myosin enhances the ATPase activity of heavy meromyosin
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Nazanin Bohlooli Ghashghaee
Abstract
Contractility of the heart muscle is a result of sliding movements between thick and thin filaments, produced by interactions between actin and myosin during the cross-bridge cycle. Activation of the myofilament is triggered by Ca2+ binding to cardiac troponin C and is regulated through an “on/off” switching process occurring in the thin filament. Beside Ca2+ regulation, strongly bound cross-bridges exert a positive feedback on myofilament regulation. Despite the importance of this positive feedback mechanism, its full molecular basis has so far remained elusive. Ca2+-regulated interactions between thick and thin filaments are widely regarded as an allosteric system, which means that multiple protein-protein interactions at their interface may exert alternative feedback effects on myofilament activation. To advance knowledge about these regulatory feedback mechanisms, we investigated a previously unstudied, hypothetical interaction between cardiac troponin and myosin, and how this interaction affects the function of myosin. Our results strongly suggest that myosin does indeed interact with the N-terminus of cardiac troponin I and the C-terminus of cardiac troponin T, suggesting a possible direct interaction between myosin and the IT-arm of troponin. We also conducted an in vitro heavy meromyosin (HMM) ATPase assay, and found that troponin significantly enhanced the actin-activated ATPase activity of HMM, both in the absence of tropomyosin and at the activated state of thin filament.
Acknowledgements
This work was supported by National Institutes of Health Grants R01 HL80186 (to W.-J.D.), and R21 HL109693 (to W.-J.D.). The M.J. Murdock Charitable Trust also provided important financial support (to W.-J.D.).
Disclosures
No conflicts of interest, financial or otherwise, are declared by authors.
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Abbreviations
- cTnC
cardiac troponin C
- cTnI
cardiac troponin I
- cTnT
cardiac troponin T
- DTT
dithiothreitol
- EGTA
ethylene glycol-bis(β-aminoethyl ether)-N,N,N’,N’-tetraacetic acid
- HMM
heavy meromyosin
- IVM
in vitro motility assay; MOPS, 3-(N-morpholino)propanesulfonic acid
- N-cTnC
N-terminus of cardiac troponin C
- Pi
inorganic phosphate
- SL
sarcomere length
- TBS/T
Tris-buffered saline solution containing 1% tween-20
- TF
thin filament
- Tm
tropomyosin
- Tn
troponin
©2017 Institute of Molecular Biology, Slovak Academy of Sciences
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- Distribution of the genus Veigaia (Mesostigmata: Veigaiidae) in Romania with notes on the species ecology
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- Chromosomal characteristics of rDNA in a conserved karyotype of two Sternopygus macrurus (Gymnotiformes: Sternopygidae) populations from upper Paraná River basin
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