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

Ksenia Terekhova, Sabine Pokutta, Yee S. Kee, Jing Li, Emad Tajkhorshid, Gerald Fuller, Alexander R. Dunn, William I. Weis

Research output: Contribution to journalArticle

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.

Original languageEnglish (US)
Article number15375
JournalScientific reports
Volume9
Issue number1
DOIs
StatePublished - Dec 1 2019

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Catenins
Cysteine
Vinculin
Actins
Cadherins
Adherens Junctions
Calorimetry
Mechanical Stress
Actin Cytoskeleton
Cell Adhesion
Mass Spectrometry

ASJC Scopus subject areas

  • General

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Binding partner- and force-promoted changes in αE-catenin conformation probed by native cysteine labeling. / Terekhova, Ksenia; Pokutta, Sabine; Kee, Yee S.; Li, Jing; Tajkhorshid, Emad; Fuller, Gerald; Dunn, Alexander R.; Weis, William I.

In: Scientific reports, Vol. 9, No. 1, 15375, 01.12.2019.

Research output: Contribution to journalArticle

Terekhova, Ksenia ; Pokutta, Sabine ; Kee, Yee S. ; Li, Jing ; Tajkhorshid, Emad ; Fuller, Gerald ; Dunn, Alexander R. ; Weis, William I. / Binding partner- and force-promoted changes in αE-catenin conformation probed by native cysteine labeling. In: Scientific reports. 2019 ; Vol. 9, No. 1.
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