© 2000 American Heart Association, Inc.
Molecular Medicine |
Single-Molecule Mechanics of R403Q Cardiac Myosin Isolated From the Mouse Model of Familial Hypertrophic Cardiomyopathy
From the Department of Molecular Physiology and Biophysics (M.J.T., E.H., D.M.W.), University of Vermont, Burlington, Vt; Howard Hughes Medical Institute and Department of Genetics (M.G., J.G.S.), Harvard Medical School, Boston, Mass; and Howard Hughes Medical Institute (C.E.S.), Brigham and Women’s Hospital, Boston, Mass. M.J.T.’s present affiliation is Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, Conn.
Correspondence to David M. Warshaw, PhD, Department of Molecular Physiology and Biophysics, Given Building, D217, University of Vermont, Burlington, VT 05405. E-mail warshaw{at}salus.med.uvm.edu
Abstract—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 ß-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 (
1 pN) and displacements (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.
Key Words: familial hypertrophic cardiomyopathy • cardiac myosin • R403Q mouse model • laser trap • molecular motor
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|
|
|
|
|
|
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