Mechanical activation of vinculin binding to talin locks talin in an unfolded conformation

Mingxi Yao; Benjamin T Goult; Hu Chen; Peiwen Cong; Michael P Sheetz; Jie Yan

doi:10.1038/srep04610

Mechanical activation of vinculin binding to talin locks talin in an unfolded conformation

Sci Rep. 2014 Apr 9:4:4610. doi: 10.1038/srep04610.

Authors

Mingxi Yao¹, Benjamin T Goult², Hu Chen³, Peiwen Cong⁴, Michael P Sheetz⁵, Jie Yan⁶

Affiliations

¹ 1] Mechanobiology Institute, National University of Singapore, 117411, Singapore [2].
² 1] Department of Biochemistry, University of Leicester, Henry Wellcome Building, Leicester, LE1 9HN, UK [2].
³ 1] Mechanobiology Institute, National University of Singapore, 117411, Singapore [2] Department of Physics, Xiamen University, Xiamen, Fujian, 361005, China.
⁴ 1] Mechanobiology Institute, National University of Singapore, 117411, Singapore [2] Singapore-MIT Alliance for Research and Technology, National University of Singapore, 117543, Singapore.
⁵ 1] Mechanobiology Institute, National University of Singapore, 117411, Singapore [2] Department of Biological Sciences, Columbia University, New York, NY 10027, USA.
⁶ 1] Mechanobiology Institute, National University of Singapore, 117411, Singapore [2] Department of Physics, National University of Singapore, 117542, Singapore [3] Centre for Bioimaging Sciences, National University of Singapore, 117546, Singapore.

Abstract

The force-dependent interaction between talin and vinculin plays a crucial role in the initiation and growth of focal adhesions. Here we use magnetic tweezers to characterise the mechano-sensitive compact N-terminal region of the talin rod, and show that the three helical bundles R1-R3 in this region unfold in three distinct steps consistent with the domains unfolding independently. Mechanical stretching of talin R1-R3 enhances its binding to vinculin and vinculin binding inhibits talin refolding after force is released. Mutations that stabilize R3 identify it as the initial mechano-sensing domain in talin, unfolding at ∼5 pN, suggesting that 5 pN is the force threshold for vinculin binding and adhesion progression.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Binding Sites
Focal Adhesions / metabolism
Magnetics
Mechanical Phenomena*
Molecular Conformation
Protein Binding
Protein Conformation
Protein Folding*
Protein Structure, Tertiary
Stress, Mechanical
Surface Plasmon Resonance
Talin / metabolism*
Vinculin / metabolism*

Substances

Talin
Vinculin

Abstract

Publication types

MeSH terms

Substances

Grants and funding