title: Cardiomyopathy mutations reveal variable region of myosin converter as major element of cross-bridge compliance.
abstract: The ability of myosin to generate motile forces is based on elastic distortion of a structural element of the actomyosin complex (cross-bridge) that allows strain to develop before filament sliding. Addressing the question, which part of the actomyosin complex experiences main elastic distortion, we suggested previously that the converter domain might be the most compliant region of the myosin head domain. Here we test this proposal by studying functional effects of naturally occurring missense mutations in the beta-myosin heavy chain, 723Arg --> Gly (R723G) and 736Ile --> Thr (I736T), in comparison to 719Arg --> Trp (R719W). All three mutations are associated with hypertrophic cardiomyopathy and are located in the converter region of the myosin head domain. We determined several mechanical parameters of single skinned slow fibers isolated from Musculus soleus biopsies of hypertrophic cardiomyopathy patients and healthy controls. Major findings of this study for mutation R723G were i), a >40% increase in fiber stiffness in rigor with a 2.9-fold increase in stiffness per myosin head (S( *)(rigor R723G) = 0.84 pN/nm S( *)(rigor WT) = 0.29 pN/nm); and ii), a significant increase in force per head (F( *)(10 degrees C), 1.99 pN vs. 1.49 pN = 1.3-fold increase; F( *)(20 degrees C), 2.56 pN vs. 1.92 pN = 1.3-fold increase) as well as stiffness per head during isometric steady-state contraction (S( *)(active10 degrees C), 0.52 pN/nm vs. 0.28 pN/nm = 1.9-fold increase). Similar changes were found for mutation R719W (2.6-fold increase in S( *)(rigor); 1.8-fold increase in F( *)(10 degrees C), 1.6-fold in F( *)(20 degrees C); twofold increase in S( *)(active10 degrees C)). Changes in active cross-bridge cycling kinetics could not account for the increase in force and active stiffness. For the above estimates the previously determined fraction of mutated myosin in the biopsies was taken into account. Data for wild-type myosin of slow soleus muscle fibers support previous findings that for the slow myosin isoform S( *) and F( *) are significantly lower than for fast myosin e.g., of rabbit psoas muscle. The data indicate that two mutations, R723G and R719W, are associated with an increase in resistance to elastic distortion of the individual mutated myosin heads whereas mutation I736T has essentially no effect. The data strongly support the notion that major elastic distortion occurs within the converter itself. Apparently, the compliance depends on specific residues, e.g., R719 and R723, presumably located at strategic positions near the long alpha-helix of the light chain binding domain. Because amino acids 719 and 723 are nonconserved residues, cross-bridge stiffness may well be specifically tuned for different myosins.
authors: Molecular and Cell Physiology, Medical School, Hannover, Germany.
authors: B
authors: B
authors: Seebohm
authors: F
authors: F
authors: Matinmehr
authors: J
authors: J
authors: Köhler
authors: A
authors: A
authors: Francino
authors: F
authors: F
authors: Navarro-Lopéz
authors: A
authors: A
authors: Perrot
authors: C
authors: C
authors: Ozcelik
authors: W J
authors: WJ
authors: McKenna
authors: B
authors: B
authors: Brenner
authors: T
authors: T
authors: Kraft
mesh_terms: D000251:Adenosine Triphosphatases / Q000378:metabolism; D024722:Cardiac Myosins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D009202:Cardiomyopathies / Q000235:genetics* / Q000503:physiopathology; D004548:Elasticity; D006801:Humans; D007537:Isometric Contraction / Q000502:physiology; D007700:Kinetics; D016014:Linear Models; D018657:Muscle Fibers, Slow-Twitch / Q000502:physiology*; D053580:Muscle Strength / Q000502:physiology; D018482:Muscle, Skeletal / Q000503:physiopathology*; D020125:Mutation, Missense; D018995:Myosin Heavy Chains / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
chemical_list: C501641:MYH7 protein, human; D000251:Adenosine Triphosphatases; D024722:Cardiac Myosins; D018995:Myosin Heavy Chains