Single-molecule mechanics of R403Q cardiac myosin isolated from the mouse model of familial hypertrophic cardiomyopathy.
Familial hypertrophic cardiomyopathy (FHC) is an inherited cardiac disease that can result in sudden death in the absence of any overt symptoms. Many of the cases documented to date have been linked with missense mutations in the beta-myosin heavy chain gene. Here we present data detailing the functional impact of one of the most deadly mutations, R403Q, on myosin motor function. Experiments were performed on whole cardiac myosin purified from a mouse model of FHC to eliminate potential uncertainties associated with protein expression systems. The R403Q mutant myosin demonstrated 2.3-fold higher actin-activated ATPase activity, 2.2-fold greater average force generation, and 1.6-fold faster actin filament sliding in the motility assay. The force- and displacement-generating capacities of both the normal and mutant myosin were also characterized at the single molecule level in the laser trap assay. Both control and mutant generated similar unitary forces ( approximately 1 pN) and displacements ( approximately 7 nm) without any differences in event durations. On the basis of the distribution of mean unitary displacements, this mutation may possibly perturb the mechanical coordination between the 2 heads of cardiac myosin. Any of these observations could, alone or possibly in combination, result in abnormal power output and potentially a stimulus for the hypertrophic response.
Tyska
M J
MJ
Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, VT 05405, USA.
Hayes
E
E
Giewat
M
M
Seidman
C E
CE
Seidman
J G
JG
Warshaw
D M
DM
D000199:Actins / Q000502:physiology; D019943:Amino Acid Substitution; D000818:Animals; D000831:Animals, Newborn; D002312:Cardiomyopathy, Hypertrophic / Q000235:genetics / Q000503:physiopathology*; D006321:Heart / Q000502:physiology / Q000503:physiopathology*; D006801:Humans; D007700:Kinetics; D051379:Mice; D008822:Mice, Transgenic; D020125:Mutation, Missense; D018995:Myosin Heavy Chains / Q000235:genetics*; D009218:Myosins / Q000378:metabolism / Q000502:physiology*; D017354:Point Mutation; D011995:Recombination, Genetic
D000199:Actins; D018995:Myosin Heavy Chains; D009218:Myosins