Divalent cations crosslink vimentin intermediate filament tail domains to regulate network mechanics

J Mol Biol. 2010 Jun 18;399(4):637-44. doi: 10.1016/j.jmb.2010.04.054. Epub 2010 May 4.


Intermediate filament networks in the cytoplasm and nucleus are critical for the mechanical integrity of metazoan cells. However, the mechanism of crosslinking in these networks and the origins of their mechanical properties are not understood. Here, we study the elastic behavior of in vitro networks of the intermediate filament protein vimentin. Rheological experiments reveal that vimentin networks stiffen with increasing concentrations of Ca(2+) and Mg(2+), showing that divalent cations act as crosslinkers. We quantitatively describe the elastic response of vimentin networks over five decades of applied stress using a theory that treats the divalent cations as crosslinkers: at low stress, the behavior is entropic in origin, and increasing stress pulls out thermal fluctuations from single filaments, giving rise to a nonlinear response; at high stress, enthalpic stretching of individual filaments significantly modifies the nonlinearity. We investigate the elastic properties of networks formed by a series of protein variants with stepwise tail truncations and find that the last 11 amino acids of the C-terminal tail domain mediate crosslinking by divalent ions. We determined the single-filament persistence length, l(P) approximately 0.5 mum, and Young's modulus, Y approximately 9 MPa; both are consistent with literature values. Our results provide insight into a crosslinking mechanism for vimentin networks and suggest that divalent ions may help regulate the cytoskeletal structure and mechanical properties of cells.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Biomechanical Phenomena
  • Cations, Divalent / pharmacology
  • Cross-Linking Reagents / pharmacology
  • Elastic Modulus
  • Humans
  • In Vitro Techniques
  • Intermediate Filaments / chemistry
  • Intermediate Filaments / drug effects
  • Intermediate Filaments / physiology
  • Intermediate Filaments / ultrastructure
  • Microscopy, Electron, Transmission
  • Molecular Sequence Data
  • Multiprotein Complexes / chemistry
  • Mutagenesis
  • Peptide Fragments / chemistry
  • Peptide Fragments / genetics
  • Peptide Fragments / physiology
  • Peptide Fragments / ultrastructure
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / ultrastructure
  • Rheology
  • Vimentin / chemistry*
  • Vimentin / genetics
  • Vimentin / physiology
  • Vimentin / ultrastructure


  • Cations, Divalent
  • Cross-Linking Reagents
  • Multiprotein Complexes
  • Peptide Fragments
  • Recombinant Proteins
  • Vimentin