title: Omecamtiv Mecarbil Enhances the Duty Ratio of Human β-Cardiac Myosin Resulting in Increased Calcium Sensitivity and Slowed Force Development in Cardiac Muscle.
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 thein vitromotility assay. The ionic strength dependence ofin vitromotility 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.
authors.affiliation: From the Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033.
authors.forename: Anja M
authors.initials: AM
authors.lastname: Swenson
authors.affiliation: From the Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033.
authors.forename: Wanjian
authors.identifier: 0000-0001-7374-6442
authors.initials: W
authors.lastname: Tang
authors.affiliation: the Department of Physiology and Division of Cardiovascular Medicine, University of Kentucky, Lexington, Kentucky 40536-0298, and.
authors.forename: Cheavar A
authors.initials: CA
authors.lastname: Blair
authors.affiliation: From the Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Fetrow
authors.affiliation: From the Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033.
authors.forename: William C
authors.initials: WC
authors.lastname: Unrath
authors.affiliation: the Department of Molecular Physiology and Biophysics, Cardiovascular Research Institute, University of Vermont, Burlington, Vermont 05405.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Previs
authors.affiliation: the Department of Physiology and Division of Cardiovascular Medicine, University of Kentucky, Lexington, Kentucky 40536-0298, and.
authors.forename: Kenneth S
authors.initials: KS
authors.lastname: Campbell
authors.affiliation: From the Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, Hershey, Pennsylvania 17033, cmy11@psu.edu.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Yengo
mesh_terms: D000199:Actins / Q000737:chemistry; D000205:Actomyosin / Q000737:chemistry; D000244:Adenosine Diphosphate / Q000737:chemistry; D000818:Animals; D002118:Calcium / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D051379:Mice; D009206:Myocardium / Q000378:metabolism*; D009218:Myosins / Q000737:chemistry; D000072417:Protein Domains; D011994:Recombinant Proteins / Q000737:chemistry; D013314:Stress, Mechanical; D014508:Urea / Q000031:analogs & derivatives* / Q000737:chemistry; D024723:Ventricular Myosins / Q000737:chemistry*
chemical_list: D000199:Actins; D011994:Recombinant Proteins; C547293:omecamtiv mecarbil; D000244:Adenosine Diphosphate; D014508:Urea; D000205:Actomyosin; D024723:Ventricular Myosins; D009218:Myosins; D002118:Calcium
keywords: actin; calcium sensitivity; cardiac muscle; contractile protein; molecular motor; myosin