Zheng, KaiLaurence, Jennifer S.Kuczera, KrzysztofVerkhivker, Gennady M.Middaugh, C. RussellSiahaan, Teruna J.2017-05-312017-05-312009-06Zheng, K., Laurence, J. S., Kuczera, K., Verkhivker, G., Russell Middaugh, C. and Siahaan, T. J. (2009), Characterization of Multiple Stable Conformers of the EC5 Domain of E-cadherin and the Interaction of EC5 with E-cadherin Peptides. Chemical Biology & Drug Design, 73: 584–598. doi:10.1111/j.1747-0285.2009.00818.xhttps://hdl.handle.net/1808/24326The objectives of this work were to express the EC5 domain of E-cadherin and determine its structural characteristics as well as to evaluate the binding properties of HAV and BLG4 peptides to EC5 using spectroscopic methods. Homophilic interactions of E-cadherins are responsible for cell-cell adhesion in the adherens junctions of the biological barriers (i.e., intestinal mucosa and blood-brain barriers). The EC5 domain of E-cadherin has an important role in T-cell adhesion to intestinal mucosa via αEβ7 integrin-E-cadherin interactions. In this study, the expressed EC5 has a high thermal stability (Tm = 64.3 °C); it also has two stable conformations at room temperature, which convert to one conformation at approximately 54.5 °C. NMR and FTIR showed that HAV and BLG4 peptides bind to EC5. HSQC-NMR showed that either Asn or Gln of EC5 was involved in the interactions with HAV and BLG4 peptides. EC5 underwent a conformational change upon interaction with the HAV and BLG4 peptides. Finally, the binding properties of both peptides were modeled by docking experiments, and the results suggest that Asn-46 and Asn-75 of EC5 could be involved during the interaction with the peptides and that the Ser and Trp residues of the HAV and BLG4 peptides, respectively, were important for binding to EC5.E-cadherinEC5 domainConformationSpectroscopyAdherens junctionCell-cell adhesionPeptide bindingArticle10.1111/j.1747-0285.2009.00818.xPMC3328966openAccess