title: Cardiac myosin isoforms exhibit differential rates of MgADP release and MgATP binding detected by myocardial viscoelasticity.
abstract: We measured myosin crossbridge detachment rate and the rates of MgADP release and MgATP binding in mouse and rat myocardial strips bearing one of the two cardiac myosin heavy chain (MyHC) isoforms. Mice and rats were fed an iodine-deficient, propylthiouracil diet resulting in ~100% expression of β-MyHC in the ventricles. Ventricles of control animals expressed ~100% α-MyHC. Chemically-skinned myocardial strips prepared from papillary muscle were subjected to sinusoidal length perturbation analysis at maximum calcium activation pCa 4.8 and 17°C. Frequency characteristics of myocardial viscoelasticity were used to calculate crossbridge detachment rate over 0.01 to 5mM [MgATP]. The rate of MgADP release, equivalent to the asymptotic value of crossbridge detachment rate at high MgATP, was highest in mouse α-MyHC (111.4±6.2s(-1)) followed by rat α-MyHC (65.0±7.3s(-1)), mouse β-MyHC (24.3±1.8s(-1)) and rat β-MyHC (15.5±0.8s(-1)). The rate of MgATP binding was highest in mouse α-MyHC (325±32 mM(-1) s(-1)) then mouse β-MyHC (152±23 mM(-1) s(-1)), rat α-MyHC (108±10 mM(-1) s(-1)) and rat β-MyHC (55±6 mM(-1) s(-1)). Because the events of MgADP release and MgATP binding occur in a post power-stroke state of the myosin crossbridge, we infer that MgATP release and MgATP binding must be regulated by isoform- and species-specific structural differences located outside the nucleotide binding pocket, which is identical in sequence for these four myosins. We postulate that differences in the stiffness profile of the entire myosin molecule, including the thick filament and the myosin-actin interface, are primarily responsible for determining the strain on the nucleotide binding pocket and the subsequent differences in the rates of nucleotide release and binding observed among the four myosins examined here.
authors.affiliation: Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, VT 05405, USA.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Wang
authors.forename: Bertrand C W
authors.initials: BC
authors.lastname: Tanner
authors.forename: Andrew T
authors.initials: AT
authors.lastname: Lombardo
authors.forename: Sarah M
authors.initials: SM
authors.lastname: Tremble
authors.forename: David W
authors.initials: DW
authors.lastname: Maughan
authors.forename: Peter
authors.initials: P
authors.lastname: Vanburen
authors.forename: Martin M
authors.initials: MM
authors.lastname: Lewinter
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Robbins
authors.forename: Bradley M
authors.initials: BM
authors.lastname: Palmer
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism*; D000469:Alkaline Phosphatase / Q000494:pharmacology; D000818:Animals; D001696:Biomechanical Phenomena; D055119:Elastic Modulus; D006352:Heart Ventricles / Q000378:metabolism*; D007037:Hypothyroidism / Q000378:metabolism; D066298:In Vitro Techniques; D007455:Iodine / Q000172:deficiency; D007700:Kinetics; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000502:physiology*; D018995:Myosin Heavy Chains / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D020033:Protein Isoforms / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D051381:Rats; D011921:Rats, Inbred WKY; D013045:Species Specificity; D019210:Troponin I / Q000378:metabolism; D014783:Viscosity
chemical_list: D020033:Protein Isoforms; D019210:Troponin I; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D007455:Iodine; D000469:Alkaline Phosphatase; D018995:Myosin Heavy Chains