Selective perturbation of the myosin recovery stroke by point mutations at the base of the lever arm affects ATP hydrolysis and phosphate release

J Biol Chem. 2007 Jun 15;282(24):17658-64. doi: 10.1074/jbc.M701447200. Epub 2007 Apr 21.

Abstract

After ATP binding the myosin head undergoes a large structural rearrangement called the recovery stroke. This transition brings catalytic residues into place to enable ATP hydrolysis, and at the same time it causes a swing of the myosin lever arm into a primed state, which is a prerequisite for the power stroke. By introducing point mutations into a subdomain interface at the base of the myosin lever arm at positions Lys(84) and Arg(704), we caused modulatory changes in the equilibrium constant of the recovery stroke, which we could accurately resolve using the fluorescence signal of single tryptophan Dictyostelium myosin II constructs. Our results shed light on a novel role of the recovery stroke: fine-tuning of this reversible equilibrium influences the functional properties of myosin through controlling the effective rates of ATP hydrolysis and phosphate release.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • Animals
  • Dictyostelium / metabolism
  • Models, Molecular
  • Myosins / chemistry
  • Myosins / genetics*
  • Myosins / metabolism*
  • Phosphates / metabolism*
  • Point Mutation*
  • Protein Structure, Tertiary*
  • Tryptophan / metabolism

Substances

  • Phosphates
  • Tryptophan
  • Adenosine Triphosphate
  • Myosins