Binding partner- and force-promoted changes in αE-catenin conformation probed by native cysteine labeling

Research output: Contribution to journalArticle

Authors

External Institution(s)

  • Stanford University
  • University of Illinois at Urbana-Champaign
  • The University of Chicago

Details

Original languageEnglish (US)
Article number15375
JournalScientific reports
Volume9
Issue number1
StatusPublished - Dec 1 2019
Peer-reviewedYes

Abstract

Adherens Junctions (AJs) are cell-cell adhesion complexes that sense and propagate mechanical forces by coupling cadherins to the actin cytoskeleton via β-catenin and the F-actin binding protein αE-catenin. When subjected to mechanical force, the cadherin•catenin complex can tightly link to F-actin through αE-catenin, and also recruits the F-actin-binding protein vinculin. In this study, labeling of native cysteines combined with mass spectrometry revealed conformational changes in αE-catenin upon binding to the E-cadherin•β-catenin complex, vinculin and F-actin. A method to apply physiologically meaningful forces in solution revealed force-induced conformational changes in αE-catenin when bound to F-actin. Comparisons of wild-type αE-catenin and a mutant with enhanced vinculin affinity using cysteine labeling and isothermal titration calorimetry provide evidence for allosteric coupling of the N-terminal β-catenin-binding and the middle (M) vinculin-binding domain of αE-catenin. Cysteine labeling also revealed possible crosstalk between the actin-binding domain and the rest of the protein. The data provide insight into how binding partners and mechanical stress can regulate the conformation of full-length αE-catenin, and identify the M domain as a key transmitter of conformational changes.

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Terekhova, K, Pokutta, S, Kee, YS, Li, J, Tajkhorshid, E, Fuller, G, Dunn, AR & Weis, WI 2019, 'Binding partner- and force-promoted changes in αE-catenin conformation probed by native cysteine labeling', Scientific reports, vol. 9, no. 1, 15375. https://doi.org/10.1038/s41598-019-51816-3