Abstract
The small molecule drug omecamtiv mecarbil (OM) specifically targets cardiac muscle myosin and is known to enhance cardiac muscle performance, yet its impact on human cardiac myosin motor function is unclear. We expressed and purified human β-cardiac myosin subfragment 1 (M2β-S1) containing a C-terminal Avi tag. We demonstrate that the maximum actin-activated ATPase activity of M2β-S1 is slowed more than 4-fold in the presence of OM, whereas the actin concentration required for half-maximal ATPase was reduced dramatically (30-fold). We find OM does not change the overall actin affinity. Transient kinetic experiments suggest that there are two kinetic pathways in the presence of OM. The dominant pathway results in a slow transition between actomyosin·ADP states and increases the time myosin is strongly bound to actin. However, OM also traps a population of myosin heads in a weak actin affinity state with slow product release. We demonstrate that OM can reduce the actin sliding velocity more than 100-fold in the in vitro motility assay. The ionic strength dependence of in vitro motility suggests the inhibition may be at least partially due to drag forces from weakly attached myosin heads. OM causes an increase in duty ratio examined in the motility assay. Experiments with permeabilized human myocardium demonstrate that OM increases calcium sensitivity and slows force development (ktr) in a concentration-dependent manner, whereas the maximally activated force is unchanged. We propose that OM increases the myosin duty ratio, which results in enhanced calcium sensitivity but slower force development in human myocardium.
Document Type
Article
Publication Date
1-12-2017
Digital Object Identifier (DOI)
https://doi.org/10.1074/jbc.M116.748780
Funding Information
This work was supported by American Heart Association Grant 14GRNT20380068 and National Institutes of Health Grant R01HL127699 (to C. M. Y.), American Heart Association Grant 15GRNT25460003 and National Institutes of Health Grant UL1TR000117 (to K. S. C.), and National Institutes of Health Grant R00HL124041 (to M. J. P.).
Related Content
This article contains supplemental Fig. S1.
Repository Citation
Swenson, Anja M.; Tang, Wanjian; Blair, Cheavar A.; Fetrow, Christopher M.; Unrath, William C.; Previs, Michael J.; Campbell, Kenneth S.; and Yengo, Christopher M., "Omecamtiv Mecarbil Enhances the Duty Ratio of Human β-Cardiac Myosin Resulting in Increased Calcium Sensitivity and Slowed Force Development in Cardiac Muscle" (2017). Physiology Faculty Publications. 118.
https://uknowledge.uky.edu/physiology_facpub/118
Supplemental Figure 1
Included in
Biochemistry, Biophysics, and Structural Biology Commons, Cardiology Commons, Physiology Commons
Notes/Citation Information
Published in The Journal of Biological Chemistry, v. 292, no. 9, p. 3768-3778.
This research was originally published in The Journal of Biological Chemistry. Anja M. Swenson, Wanjian Tang, Cheavar A. Blair, Christopher M. Fetrow, William C. Unrath, Michael J. Previs, Kenneth S. Campbell, and Christopher M. Yengo. Omecamtiv Mecarbil Enhances the Duty Ratio of Human β-Cardiac Myosin Resulting in Increased Calcium Sensitivity and Slowed Force Development in Cardiac Muscle. J. Biol. Chem. 2017; 292:3768-3778. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.
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