The two heads of smooth muscle myosin are enzymatically independent but mechanically interactive.
The interaction between the two heads of myosin II during motion and force production is poorly understood. To examine this issue, we developed an expression and purification strategy to isolate homogeneous populations of heterodimeric smooth muscle heavy meromyosins containing heads with different properties. As an extreme example, we characterized a heterodimer containing one native head and one head locked in a "weak binding" state by a point mutation in switch 2 (E470A). The in vitro actin filament motility of this heterodimer was the same as the homodimeric control with two cycling heads, suggesting that only one head of a pair actively interacts with actin to generate maximal velocity. A second naturally occurring heterodimer contained two cycling heads with 2-fold different activity, due to the presence or absence of a 7-amino acid insert near the active site. Enzymatically this (+/-) insert heterodimer was indistinguishable from a (50:50) mixture of the two homodimers, but its motility averaged 17% less than that of the mixture. These data suggest that one head of a heterodimer can disproportionately affect the mechanics of double-headed myosin, a finding relevant to our understanding of heterozygous mutant myosins found in disease states like familial hypertrophic cardiomyopathy.
Rovner
Arthur S
AS
Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, Vermont 05405, USA. rovner@physiology.med.uvm.edu
Fagnant
Patricia M
PM
Trybus
Kathleen M
KM
D000244:Adenosine Diphosphate / Q000378:metabolism; D000818:Animals; D001696:Biomechanical Phenomena; D024741:Cardiomyopathy, Hypertrophic, Familial / Q000235:genetics / Q000503:physiopathology; D002645:Chickens; D019281:Dimerization; D006801:Humans; D066298:In Vitro Techniques; D015879:Myosin Subfragments / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*; D017354:Point Mutation; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D024745:Smooth Muscle Myosins / Q000737:chemistry* / Q000235:genetics / Q000502:physiology*
D015879:Myosin Subfragments; D011994:Recombinant Proteins; D000244:Adenosine Diphosphate; D024745:Smooth Muscle Myosins