Cingulin interacts with F-actin in vitro

FEBS Lett. 2001 Oct 19;507(1):21-4. doi: 10.1016/s0014-5793(01)02936-2.

Abstract

Cingulin, a M(r) 140-160 kDa protein of the cytoplasmic plaque of epithelial tight junctions (TJ), interacts in vitro with TJ proteins and myosin. Here we investigated cingulin interaction with actin, using His-tagged, full-length Xenopus laevis cingulin expressed in insect cells, and glutathione S-transferase (GST) fusion proteins of fragments of cingulin expressed in bacteria. Purified full-length cingulin co-pelleted with F-actin after high speed centrifugation, and promoted the sedimentation of F-actin under low speed centrifugation, suggesting that cingulin is an actin-cross-linking protein. The actin interaction of GST fusion proteins containing fragments of Xenopus cingulin suggested that the F-actin binding site is between residues 101 and 294.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / metabolism*
  • Animals
  • Binding Sites
  • Cross-Linking Reagents
  • Cytoskeleton / metabolism
  • Humans
  • In Vitro Techniques
  • Membrane Proteins / chemistry
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism*
  • Microfilament Proteins
  • Peptide Mapping
  • Protein Binding
  • Rabbits
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Spodoptera
  • Tight Junctions / chemistry
  • Tight Junctions / metabolism
  • Xenopus Proteins*
  • Xenopus laevis

Substances

  • Actins
  • CGN protein, Xenopus
  • CGN protein, human
  • Cross-Linking Reagents
  • Membrane Proteins
  • Microfilament Proteins
  • Recombinant Fusion Proteins
  • Xenopus Proteins