Purification and properties of transgelin: a transformation and shape change sensitive actin-gelling protein

J Cell Biol. 1993 Jun;121(5):1065-73. doi: 10.1083/jcb.121.5.1065.

Abstract

We have purified the transformation and shape change sensitive isoform of an actin associated polypeptide doublet previously described by us (Shapland, C., P. Lowings, and D. Lawson. 1988. J. Cell Biol. 107:153-161) and have shown that it is evolutionarily conserved as far back as yeast. The purified protein: (a) binds directly to actin filaments at a ratio of 1:6 actin monomers, with a binding constant (Ka) of approximately 7.5 x 10(5) M-1; and (b) causes actin filament gelation within 2 min. Although these activities are controlled by ionic strength (and may be mediated by positively charged amino acid residues) the molecule remains as a monomer irrespective of ionic conditions. EM reveals that the addition of this protein to actin filaments converts them from a loose, random distribution into a tangled, cross-linked meshwork within 1 min, and discrete tightly aggregated foci after 10 min. By use of an "add-back" cell permeabilization system we can rebind this molecule specifically to actin filaments in cells from which it has previously been removed. Since the protein is transformation sensitive and gels actin, we have named it transgelin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Cell Transformation, Neoplastic / ultrastructure
  • Gels
  • Light
  • Microfilament Proteins / chemistry
  • Microfilament Proteins / immunology
  • Microfilament Proteins / isolation & purification*
  • Molecular Sequence Data
  • Molecular Weight
  • Muscle Proteins / chemistry
  • Muscle Proteins / immunology
  • Muscle Proteins / isolation & purification*
  • Scattering, Radiation
  • Schizosaccharomyces / chemistry
  • Sheep
  • Viscosity

Substances

  • Gels
  • Microfilament Proteins
  • Muscle Proteins
  • Tagln protein, mouse
  • transgelin