Smooth, cardiac and skeletal muscle myosin force and motion generation assessed by cross-bridge mechanical interactions in vitro.
Differences in the mechanical properties of mammalian smooth, skeletal, and cardiac muscle have led to the proposal that the myosin isozymes expressed by these tissues may differ in their molecular mechanics. To test this hypothesis, mixtures of fast skeletal, V1 cardiac, V3 cardiac and smooth muscle (phosphorylated and unphosphorylated) myosin were studied in an in vitro motility assay in which fluorescently-labelled actin filaments are observed moving over a myosin coated surface. Pure populations of each myosin produced actin filament velocities proportional to their actin-activated ATPase rates. Mixtures of two myosin species produced actin filament velocities between those of the faster and slower myosin alone. However, the shapes of the myosin mixture curves depended upon the types of myosins present. Analysis of myosin mixtures data suggest that: (1) the two myosins in the mixture interact mechanically and (2) the same force-velocity relationship describes a myosin's ability to operate over both positive and negative forces. These data also allow us to rank order the myosins by their average force per cross-bridge and ability to resist motion (phosphorylated smooth > skeletal = V3 cardiac > V1 cardiac). The results of our study may reflect the mechanical consequence of multiple myosin isozyme expression in a single muscle cell.
Harris
D E
DE
University of Vermont, Department of Molecular Physiology & Biophysics, Burlington 05405.
Work
S S
SS
Wright
R K
RK
Alpert
N R
NR
Warshaw
D M
DM
D008841:Actin Cytoskeleton / Q000502:physiology; D000199:Actins / Q000502:physiology; D000205:Actomyosin / Q000502:physiology; D000818:Animals; D001696:Biomechanical Phenomena; D002645:Chickens / Q000378:metabolism; D006321:Heart / Q000502:physiology*; D007527:Isoenzymes / Q000502:physiology*; D009038:Motion; D009130:Muscle, Smooth / Q000201:enzymology / Q000502:physiology*; D009132:Muscles / Q000201:enzymology / Q000502:physiology*; D009206:Myocardium / Q000201:enzymology; D009218:Myosins / Q000302:isolation & purification / Q000502:physiology*; D010766:Phosphorylation; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D013329:Structure-Activity Relationship; D014422:Turkeys / Q000378:metabolism
D000199:Actins; D007527:Isoenzymes; D000205:Actomyosin; D009218:Myosins