Biochemical Journal

Research article

Molecular interactions between desmosomal cadherins

Shabih-e-Hassnain SYED, Brian TRINNAMAN, Stephen MARTIN, Sarah MAJOR, Jon HUTCHINSON, Anthony I. MAGEE

Abstract

Desmocollins (Dscs) and desmogleins (Dsgs) are cell-adhesion molecules involved in the formation of desmosome cell—cell junctions and share structural similarities to classical cadherins such as E-cadherin. In order to identify and provide quantitative information on the types of protein—protein interactions displayed by the type 2 isoforms and investigate the role of Ca2+ in this process, we have developed an Escherichia coli expression system to generate recombinant proteins containing the first two extracellular domains, namely Dsg2(1-2) and Dsc2(1-2). Analytical ultracentrifugation, chemical cross-linking, CD, fluorescence and BIAcore have been used to provide the first direct evidence of Ca2+ binding to desmosomal cadherins. These studies suggest that Dsc2(1-2) not only exhibits homophilic interactions in solution, but can also form heterophilic interactions with Dsg2(1-2). The latter, on the other hand, shows much weaker homophilic association. Our results further demonstrate that heterophilic interactions are Ca2+-dependent, whereas the Ca2+-dependence of homophilic association is less clear. Our data indicate that the functional properties of Dsc2(1-2) are more similar to those of classical cadherins, consistent with the observation that Dsc shares a higher level of sequence homology with classical cadherins than does Dsg. In addition to corroborating the conclusions of previously reported transfection studies which suggest the formation of lateral heterodimers and homodimers, our results also provide direct quantitative information on the strength of these interactions which are essential for understanding the adhesion mechanism.

  • Ca2+ binding
  • cell adhesion
  • desmocollin
  • desmoglein
  • dimerization

Footnotes

  • Abbreviations used: Dsg, desmoglein; Dsc, desmocollin; NC1, first domain of N-cadherin; Ecad1-2, the two N-terminal domains of E-cadherin; Dsg2(1-2), the two N-terminal domains of desmoglein2; Dsc2(1-2), the two N-terminal domains of desmocollin 2; GST, glutathione S-transferase; DTT, dithiothreitol; IPTG, isopropyl β-d-thiogalactopyranoside; Caps, 3-(cyclohexylamino)propane-1-sulphonic acid; CAMs, cell-adhesion molecules; FXa, Factor Xa (or activated Factor X), a restriction protease from Roche; Ni-NTA—agarose, Ni2+-nitrilotriacetate—agarose; BS3, bis(sulphosuccinimidyl) suberate.