Evidence for a conformational change in actin induced by fimbrin (N375) binding

J Cell Biol. 1997 Oct 20;139(2):387-96. doi: 10.1083/jcb.139.2.387.

Abstract

Fimbrin belongs to a superfamily of actin cross-linking proteins that share a conserved 27-kD actin-binding domain. This domain contains a tandem duplication of a sequence that is homologous to calponin. Calponin homology (CH) domains not only cross-link actin filaments into bundles and networks, but they also bind intermediate filaments and some signal transduction proteins to the actin cytoskeleton. This fundamental role of CH domains as a widely used actin-binding domain underlines the necessity to understand their structural interaction with actin. Using electron cryomicroscopy, we have determined the three-dimensional structure of F-actin and F-actin decorated with the NH2-terminal CH domains of fimbrin (N375). In a difference map between actin filaments and N375-decorated actin, one end of N375 is bound to a concave surface formed between actin subdomains 1 and 2 on two neighboring actin monomers. In addition, a fit of the atomic model for the actin filament to the maps reveals the actin residues that line, the binding surface. The binding of N375 changes actin, which we interpret as a movement of subdomain 1 away from the bound N375. This change in actin structure may affect its affinity for other actin-binding proteins and may be part of the regulation of the cytoskeleton itself. Difference maps between actin and actin decorated with other proteins provides a way to look for novel structural changes in actin.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Actins / chemistry
  • Actins / metabolism*
  • Actins / ultrastructure*
  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Calcium-Binding Proteins / chemistry
  • Calponins
  • Cloning, Molecular
  • Computer Simulation
  • Conserved Sequence
  • Humans
  • Membrane Glycoproteins / chemistry
  • Membrane Glycoproteins / metabolism*
  • Membrane Glycoproteins / ultrastructure*
  • Microfilament Proteins / chemistry
  • Microfilament Proteins / metabolism
  • Microscopy, Electron
  • Models, Molecular
  • Models, Structural
  • Protein Binding
  • Protein Conformation*
  • Rabbits
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism

Substances

  • Actins
  • Calcium-Binding Proteins
  • Membrane Glycoproteins
  • Microfilament Proteins
  • Recombinant Proteins
  • plastin