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J Mol Biol. 2007 Aug 24;371(4):902-13. Epub 2007 Jun 2.

Evidence for an interaction between the SH3 domain and the N-terminal extension of the essential light chain in class II myosins.

Author information

1
Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, VT 05405, USA. lowey@physiology.med.uvm.edu

Abstract

The function of the src-homology 3 (SH3) domain in class II myosins, a distinct beta-barrel structure, remains unknown. Here, we provide evidence, using electron cryomicroscopy, in conjunction with light-scattering, fluorescence and kinetic analyses, that the SH3 domain facilitates the binding of the N-terminal extension of the essential light chain isoform (ELC-1) to actin. The 41 residue extension contains four conserved lysine residues followed by a repeating sequence of seven Pro/Ala residues. It is widely believed that the highly charged region interacts with actin, while the Pro/Ala-rich sequence forms a rigid tether that bridges the approximately 9 nm distance between the myosin lever arm and the thin filament. In order to localize the N terminus of ELC in the actomyosin complex, an engineered Cys was reacted with undecagold-maleimide, and the labeled ELC was exchanged into myosin subfragment-1 (S1). Electron cryomicroscopy of S1-bound actin filaments, together with computer-based docking of the skeletal S1 crystal structure into 3D reconstructions, showed a well-defined peak for the gold cluster near the SH3 domain. Given that SH3 domains are known to bind proline-rich ligands, we suggest that the N-terminal extension of ELC interacts with actin and modulates myosin kinetics by binding to the SH3 domain during the ATPase cycle.

PMID:
17597155
PMCID:
PMC2693010
DOI:
10.1016/j.jmb.2007.05.080
[Indexed for MEDLINE]
Free PMC Article

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