Adhesive and lateral E-cadherin dimers are mediated by the same interface.
E-cadherin is a transmembrane protein that mediates Ca(2+)-dependent cell-cell adhesion. To study cadherin-cadherin interactions that may underlie the adhesive process, a recombinant E-cadherin lacking free sulfhydryl groups and its mutants with novel cysteines were expressed in epithelial A-431 cells. These cysteine mutants, designed according to various structural models of cadherin dimers, were constructed to reveal cadherin dimerization by the bifunctional sulfhydryl-specific cross-linker BM[PE0]3. Cross-linking experiments with the mutants containing a cysteine at strand B of their EC1 domains did show cadherin dimerization. By their properties these dimers correspond to those which have been characterized by co-immunoprecipitation assay. Under standard culture conditions the adhesive dimer is a dominant form. Calcium depletion dissociates adhesive dimers and promotes the formation of lateral dimers. Our data show that both dimers are mediated by the amino-terminal cadherin domain. Furthermore, the interfaces involved in both adhesive and lateral dimerization appear to be the same. The coexistence of the structurally identical adhesive and lateral dimers suggests some flexibility of the extracellular cadherin region.
Division of Dermatology, Washington University Medical School, 660 S. Euclid Avenue, St. Louis, MO 63110, USA.
Regina B
RB
Troyanovsky
Eugene
E
Sokolov
Sergey M
SM
Troyanovsky
D001665:Binding Sites; D015820:Cadherins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology; D002460:Cell Line; D003432:Cross-Linking Reagents; D019281:Dimerization; D006801:Humans; D066298:In Vitro Techniques; D008958:Models, Molecular; D016296:Mutagenesis; D020836:Protein Structure, Quaternary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D015820:Cadherins; D003432:Cross-Linking Reagents; D011994:Recombinant Proteins; D002118:Calcium