Understanding the role of the G-actin-binding domain of Ena/VASP in actin assembly

J Struct Biol. 2006 Aug;155(2):195-201. doi: 10.1016/j.jsb.2006.01.012. Epub 2006 Apr 25.

Abstract

The Ena/VASP and WASP family of proteins play distinct roles in actin cytoskeleton remodeling. Ena/VASP is linked to actin filament elongation, whereas WASP plays a role in filament nucleation and branching mediated by Arp2/3 complex. The molecular mechanisms controlling both processes are only emerging. Both Ena/VASP and WASP are multidomain proteins. They both present poly-Pro regions, which mediate the binding of profilin-actin, followed by G-actin-binding (GAB) domains of the WASP-homology 2 (WH2) type. However, the WH2 of Ena/VASP is somewhat different from that of WASP, and has been poorly characterized. Here we demonstrate that this WH2 binds profilin-actin with higher affinity than actin alone. The results are consistent with a model whereby allosteric modulation of affinity drives the transition of profilin-actin from the poly-Pro region to the WH2 and then to the barbed end of the filament during elongation. Therefore, the function of the WH2 in Ena/VASP appears to be to "process" profilin-actin for its incorporation at the barbed end of the growing filament. Conformational changes in the newly incorporated actin subunit, resulting either from nucleotide hydrolysis or from the G- to F-actin transition, may serve as a "sensor" for the processive stepping of Ena/VASP. Conserved domain architecture suggests that WASP may work similarly.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Actin Cytoskeleton / chemistry
  • Actin Cytoskeleton / metabolism
  • Actins / chemistry
  • Actins / genetics
  • Actins / metabolism*
  • Amino Acid Sequence
  • Animals
  • Calorimetry
  • Cell Adhesion Molecules / chemistry
  • Cell Adhesion Molecules / genetics
  • Cell Adhesion Molecules / metabolism*
  • Humans
  • Kinetics
  • Microfilament Proteins / chemistry
  • Microfilament Proteins / genetics
  • Microfilament Proteins / metabolism*
  • Models, Molecular
  • Molecular Sequence Data
  • Phosphoproteins / chemistry
  • Phosphoproteins / genetics
  • Phosphoproteins / metabolism*
  • Profilins / chemistry
  • Profilins / metabolism
  • Protein Binding
  • Protein Structure, Tertiary
  • Rabbits
  • Sequence Homology, Amino Acid
  • Structure-Activity Relationship
  • Wiskott-Aldrich Syndrome Protein / chemistry
  • Wiskott-Aldrich Syndrome Protein / genetics
  • Wiskott-Aldrich Syndrome Protein / metabolism*

Substances

  • Actins
  • Cell Adhesion Molecules
  • Microfilament Proteins
  • Phosphoproteins
  • Profilins
  • Wiskott-Aldrich Syndrome Protein
  • vasodilator-stimulated phosphoprotein