Cardiac myosin activation with 2-deoxy-ATP via increased electrostatic interactions with actin.

Autor:	Powers JD; Department of Bioengineering, University of Washington, Seattle, WA 98109; j2powers@ucsd.edu mregnier@uw.edu.; Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093., Yuan CC; Department of Bioengineering, University of Washington, Seattle, WA 98109., McCabe KJ; Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093., Murray JD; Department of Bioengineering, University of Washington, Seattle, WA 98109.; Department of Physiology and Biophysics, University of Washington, Seattle, WA 98109., Childers MC; Department of Bioengineering, University of Washington, Seattle, WA 98109., Flint GV; Department of Bioengineering, University of Washington, Seattle, WA 98109., Moussavi-Harami F; Division of Cardiology, Department of Medicine, University of Washington, Seattle, WA 98109., Mohran S; Department of Bioengineering, University of Washington, Seattle, WA 98109., Castillo R; Department of Bioengineering, University of Washington, Seattle, WA 98109., Zuzek C; Department of Bioengineering, University of Washington, Seattle, WA 98109., Ma W; Department of Biological Sciences, Illinois Institute of Technology, Chicago, IL 60616., Daggett V; Department of Bioengineering, University of Washington, Seattle, WA 98109., McCulloch AD; Department of Bioengineering, University of California, San Diego, La Jolla, CA 92093., Irving TC; Department of Biological Sciences, Illinois Institute of Technology, Chicago, IL 60616., Regnier M; Department of Bioengineering, University of Washington, Seattle, WA 98109; j2powers@ucsd.edu mregnier@uw.edu.; Department of Physiology and Biophysics, University of Washington, Seattle, WA 98109.
Jazyk:	angličtina
Zdroj:	Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 2019 Jun 04; Vol. 116 (23), pp. 11502-11507. Date of Electronic Publication: 2019 May 20.
DOI:	10.1073/pnas.1905028116
Abstrakt:	The naturally occurring nucleotide 2-deoxy-adenosine 5'-triphosphate (dATP) can be used by cardiac muscle as an alternative energy substrate for myosin chemomechanical activity. We and others have previously shown that dATP increases contractile force in normal hearts and models of depressed systolic function, but the structural basis of these effects has remained unresolved. In this work, we combine multiple techniques to provide structural and functional information at the angstrom-nanometer and millisecond time scales, demonstrating the ability to make both structural measurements and quantitative kinetic estimates of weak actin-myosin interactions that underpin sarcomere dynamics. Exploiting dATP as a molecular probe, we assess how small changes in myosin structure translate to electrostatic-based changes in sarcomere function to augment contractility in cardiac muscle. Through Brownian dynamics simulation and computational structural analysis, we found that deoxy-hydrolysis products [2-deoxy-adenosine 5'-diphosphate (dADP) and inorganic phosphate (Pi)] bound to prepowerstroke myosin induce an allosteric restructuring of the actin-binding surface on myosin to increase the rate of cross-bridge formation. We then show experimentally that this predicted effect translates into increased electrostatic interactions between actin and cardiac myosin in vitro. Finally, using small-angle X-ray diffraction analysis of sarcomere structure, we demonstrate that the proposed increased electrostatic affinity of myosin for actin causes a disruption of the resting conformation of myosin motors, resulting in their repositioning toward the thin filament before activation. The dATP-mediated structural alterations in myosin reported here may provide insight into an improved criterion for the design or selection of small molecules to be developed as therapeutic agents to treat systolic dysfunction. Competing Interests: Conflict of interest statement: M.R. holds an international patent (US Patent 9,868,937 B2) on cell and gene-based methods to improve cardiac function by elevating 2′-deoxy-adenosine 5′-triphosphate. A.D.M. is a cofounder of and has an equity interest in Insilicomed, Inc. and Vektor Medical, Inc., and he serves on the scientific advisory boards. Some of his research grants acknowledged here have been identified for conflict of interest management based on the overall scope of the project and its potential benefit to these entities. The author is required by his institution to disclose this relationship in publications acknowledging the grant support. However, the research subject and findings reported here did not involve the companies in any way and have no relationship whatsoever to the business activities or scientific interests of the companies. The terms of this arrangement have been reviewed and approved by the University of California, San Diego in accordance with its conflict of interest policies. The other authors have no competing interests to declare.
Databáze:	MEDLINE
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