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Structural basis for vinculin activation at sites of cell adhesion

Nature volume 430, pages 583–586 (2004)Cite this article

Abstract

Vinculin is a highly conserved intracellular protein with a crucial role in the maintenance and regulation of cell adhesion and migration1,2,3. In the cytosol, vinculin adopts a default autoinhibited conformation4,5. On recruitment to cell–cell and cell–matrix adherens-type junctions, vinculin becomes activated and mediates various protein–protein interactions that regulate the links between F-actin and the cadherin and integrin families of cell-adhesion molecules. Here we describe the crystal structure of the full-length vinculin molecule (1,066 amino acids), which shows a five-domain autoinhibited conformation in which the carboxy-terminal tail domain is held pincer-like by the vinculin head, and ligand binding is regulated both sterically and allosterically. We show that conformational changes in the head, tail and proline-rich domains are linked structurally and thermodynamically, and propose a combinatorial pathway to activation that ensures that vinculin is activated only at sites of cell adhesion when two or more of its binding partners are brought into apposition.

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Figure 1: Structure of full-length vinculin in its autoinhibited state.
Figure 2: Accessible surface of Vt in the context of the full-length molecule.
Figure 3: Comparison of vinculin domains 3b and 4 with the ‘M fragment’ of α-catenin.
Figure 4: Calorimetric analysis of vinculin and ligand complexes.

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Acknowledgements

We thank the Cell Migration Consortium, the NIH, the BBSRC and the Wellcome Trust for financial support; the DOE and staff at SSRL for synchrotron access and support; S. Lam for the talin rod expression vector; M. Zhang for talin expression; and B. Patel for vinculin purification. D.M.C. is supported by a Howard Hughes predoctoral fellowship.

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Authors and Affiliations

  1. Program on Cell Adhesion, The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, California, 92037, USA

    Constantina Bakolitsa, Laurie A. Bankston, Andrey A. Bobkov, Gregory W. Cadwell & Robert C. Liddington

  2. Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland, 21205, USA

    Daniel M. Cohen & Susan W. Craig

  3. Department of Biochemistry, University of Leicester, LE1 7RH, Leicester, UK

    Lisa Jennings & David R. Critchley

Authors
  1. Constantina Bakolitsa
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  7. David R. Critchley
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Corresponding author

Correspondence to Robert C. Liddington.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure S1

Shows an alignment of vinculin and α-catenin sequences with the secondary structural elements for vinculin. (DOC 52 kb)

Supplementary Figure S2

Shows an analysis of the effects of mutations in basic collar, C-terminus and basic collar of the vinculin tail domain on PIP2 binding. (DOC 94 kb)

Supplementary Table S1

Table of thermodynamic binding parameters for the binding of -catenin D3 and a helical peptide derived from talin (VBS3 = 1944-1969) to vinculin VH. (DOC 19 kb)

Supplementary Table S2

This table summarizes the crystallographic data collection, phase determination and refinement. (DOC 26 kb)

Supplementary Methods

Describes methods pertaining to the data in Supplementary Figure S2, and additional notes on the method of Differential Scanning Calorimetry. (DOC 26 kb)

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Bakolitsa, C., Cohen, D., Bankston, L. et al. Structural basis for vinculin activation at sites of cell adhesion. Nature 430, 583–586 (2004). https://doi.org/10.1038/nature02610

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