Identification of a novel recognition sequence for fibronectin within the NH2-terminal β-sandwich domain of tissue transglutaminase Article

Hang, J, Zemskov, EA, Lorand, L et al. (2005). Identification of a novel recognition sequence for fibronectin within the NH2-terminal β-sandwich domain of tissue transglutaminase . JOURNAL OF BIOLOGICAL CHEMISTRY, 280(25), 23675-23683. 10.1074/jbc.M503323200

cited authors

  • Hang, J; Zemskov, EA; Lorand, L; Belkin, AM

authors

abstract

  • Tissue transglutaminase belongs to the multigene transglutaminase family of Ca2+-dependent protein cross-linking enzymes. Unlike other transglutaminases, it is involved in cell-matrix interactions and serves as an adhesion co-receptor for fibronectin. Previous work established that the fibronectin-binding motif(s) is located within the NH2-terminal proteolytic fragment of the protein consisting of residues 1-272. Here we identify a novel fibronectin recognition site within this sequence of tissue transglutaminase. Substitution of individual domains of tissue transglutaminase with those from homologous factor XIIIA showed that the major fibronectin-binding site is present within the first β-sandwich domain of the protein. Experiments with deletion mutants of the first domain revealed that amino acids 81-140 of tissue transglutaminase are involved in fibronectin binding. Using synthetic peptides encompassing this region, we found that the peptide 88WTATWDQQDCTLSLQLTT106 inhibited the interaction of tissue transglutaminase with fibronectin and decreased transglutaminase- dependent cell adhesion and spreading. In the three-dimensional structure of the first domain, amino acids 88-106 comprise an extended hairpin formed by antiparallel β strands 5 and 6. Mutations of Asp94 and Asp 97 within the β5/β6 hairpin to Ala significantly reduced the affinity of tissue transglutaminase for fibronectin, indicating that these residues are critical for fibronectin binding. Identification of the fibronectin-binding site on tissue transglutaminase will help to dissect the role of this protein in cell-matrix interactions. © 2005 by The American Society for Biochemistry and Molecular Biology, Inc.

publication date

  • June 24, 2005

published in

Digital Object Identifier (DOI)

start page

  • 23675

end page

  • 23683

volume

  • 280

issue

  • 25