Transglutaminase-catalyzed cross-linking of fibrils of collagen V/XI in A204 rhabdomyosarcoma cells

Biochemistry. 1995 Oct 24;34(42):13768-75. doi: 10.1021/bi00042a007.


Collagens V and XI are thought to form a core around which the major interstitial collagens, I and II, respectively, are organized during fibrillogenesis. We previously reported the presence of a heterotypic form of collagens V and XI, [alpha 1(XI)]2 alpha 2(V), in cultures of A204 rhabdomyosarcoma cells [Kleman, J.-P., Hartmann, D. J., Ramirez, F., & van der Rest, M. (1992) Eur. J. Biochem. 210, 329-335]. This collagen forms a matrix which remains highly insoluble, even when cells were cultured in the presence of beta-aminopropionitrile, an inhibitor of lysyl oxidase and thereby of "classical" collagen cross-linking. When the cells were cultured in the presence of putrescine, a competitive inhibitor of transglutaminase-catalyzed protein cross-linking, a drastic increase in collagen solubility was observed. This result indicates that a transglutaminase contributes to the covalent stabilization of the collagen matrix of these cells. A204 rhabdomyosarcoma cells express tissue transglutaminase as revealed by specific antibodies, and enzyme activity was detected in the cell layer during culture and in cell extracts. Both collagens V and XI are specific glutaminyl substrates for tissue transglutaminase in vitro, as shown by incorporation of [3H]putrescine. The highly homologous alpha 1 chains of collagens V and XI were the major targets for the cross-linking. Trypsin cleaved the [3H] label from the alpha 1 chain of collagen V, demonstrating that the cross-linking occurs in the non triple helical propeptide domains.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Aminopropionitrile / pharmacology
  • Cadaverine / analogs & derivatives
  • Cadaverine / metabolism
  • Caseins / metabolism
  • Cell Extracts
  • Collagen / chemistry
  • Collagen / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Fluorescent Antibody Technique
  • Humans
  • Immunoblotting
  • Kinetics
  • Molecular Sequence Data
  • Protein Structure, Secondary
  • Protein-Lysine 6-Oxidase / antagonists & inhibitors
  • Protein-Lysine 6-Oxidase / metabolism
  • Putrescine / metabolism
  • Putrescine / pharmacology
  • Rhabdomyosarcoma
  • Solubility
  • Transglutaminases / antagonists & inhibitors
  • Transglutaminases / metabolism*
  • Trypsin / metabolism
  • Tumor Cells, Cultured


  • Caseins
  • Cell Extracts
  • N,N-dimethylcasein
  • Aminopropionitrile
  • Collagen
  • Protein-Lysine 6-Oxidase
  • Transglutaminases
  • Trypsin
  • monodansylcadaverine
  • Cadaverine
  • Putrescine