The predicted coiled-coil domain of myosin 10 forms a novel elongated domain that lengthens the head

J Biol Chem. 2005 Oct 14;280(41):34702-8. doi: 10.1074/jbc.M504887200. Epub 2005 Jul 18.

Abstract

Myosin 10 contains a region of predicted coiled coil 120 residues long. However, the highly charged nature and pattern of charges in the proximal 36 residues appear incompatible with coiled-coil formation. Circular dichroism, NMR, and analytical ultracentrifugation show that a synthesized peptide containing this region forms a stable single alpha-helix (SAH) domain in solution and does not dimerize to form a coiled coil even at millimolar concentrations. Additionally, electron microscopy of a recombinant myosin 10 containing the motor, the three calmodulin binding domains, and the full-length predicted coiled coil showed that it was mostly monomeric at physiological protein concentration. In dimers the molecules were joined only at their extreme distal ends, and no coiled-coil tail was visible. Furthermore, the neck lengths of both monomers and dimers were much longer than expected from the number of calmodulin binding domains. In contrast, micrographs of myosin 5 heavy meromyosin obtained under the same conditions clearly showed a coiled-coil tail, and the necks were the predicted length. Thus the predicted coiled coil of myosin 10 forms a novel elongated structure in which the proximal region is a SAH domain and the distal region is a SAH domain (or has an unknown extended structure) that dimerizes only at its end. Sequence comparisons show that similar structures may exist in the predicted coiled-coil domains of myosins 6 and 7a and MyoM and could function to increase the size of the working stroke.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Calmodulin / chemistry
  • Cattle
  • Circular Dichroism
  • Dimerization
  • Magnetic Resonance Spectroscopy
  • Microscopy, Electron
  • Models, Molecular
  • Molecular Sequence Data
  • Myosins / chemistry*
  • Peptides / chemistry
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Sequence Homology, Amino Acid
  • Sodium Chloride / pharmacology
  • Temperature
  • Ultracentrifugation
  • Ultraviolet Rays

Substances

  • Calmodulin
  • Myo10 protein, mouse
  • Peptides
  • Recombinant Proteins
  • Sodium Chloride
  • Myosins