Cooperative vinculin binding to talin mapped by time-resolved super resolution microscopy

Xian Hu, Chaoran Jing, Xiaochun Xu, Naotaka Nakazawa, Virginia W. Cornish, Felix M. Margadant, Michael Sheetz

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The dimeric focal adhesion protein talin contains up to 22 cryptic vinculin binding sites that are exposed by unfolding. Using a novel method to monitor the in situ dynamics of the talin dimer stretch, we find that in contrast to several prevalent talin dimer models the integrin-binding talin N-termini are separated by 162 ± 44 nm on average whereas as expected the C-terminal dimerization domains colocalize and are mobile. Using vinculin tagged by DHFR-TMP Atto655 label, we found that optimal vinculin and vinculin head binding occurred when talin was stretched to 180 nm, while the controls did not bind to talin. Surprisingly, multiple vinculins bound within a single second in narrowly localized regions of the talin rod during stretching. We suggest that talin stretches as an antiparallel dimer and that activates vinculin binding in a cooperative manner, consistent with the stabilization of folded talin by other binding proteins.

Original languageEnglish (US)
Pages (from-to)4062-4068
Number of pages7
JournalNano Letters
Volume16
Issue number7
DOIs
StatePublished - Jul 13 2016
Externally publishedYes

Fingerprint

Talin
Vinculin
Dimers
Microscopic examination
microscopy
dimers
Dimerization
Binding sites
proteins
Stretching
Labels
Adhesion
Stabilization
dimerization
Proteins
adhesion
rods
stabilization
Thymidine Monophosphate
Integrins

Keywords

  • focal adhesion
  • localization microscopy
  • protein recruitment
  • single molecule tracking
  • Talin
  • vinculin

ASJC Scopus subject areas

  • Bioengineering
  • Chemistry(all)
  • Materials Science(all)
  • Condensed Matter Physics
  • Mechanical Engineering

Cite this

Cooperative vinculin binding to talin mapped by time-resolved super resolution microscopy. / Hu, Xian; Jing, Chaoran; Xu, Xiaochun; Nakazawa, Naotaka; Cornish, Virginia W.; Margadant, Felix M.; Sheetz, Michael.

In: Nano Letters, Vol. 16, No. 7, 13.07.2016, p. 4062-4068.

Research output: Contribution to journalArticle

Hu, X, Jing, C, Xu, X, Nakazawa, N, Cornish, VW, Margadant, FM & Sheetz, M 2016, 'Cooperative vinculin binding to talin mapped by time-resolved super resolution microscopy', Nano Letters, vol. 16, no. 7, pp. 4062-4068. https://doi.org/10.1021/acs.nanolett.6b00650
Hu X, Jing C, Xu X, Nakazawa N, Cornish VW, Margadant FM et al. Cooperative vinculin binding to talin mapped by time-resolved super resolution microscopy. Nano Letters. 2016 Jul 13;16(7):4062-4068. https://doi.org/10.1021/acs.nanolett.6b00650
Hu, Xian ; Jing, Chaoran ; Xu, Xiaochun ; Nakazawa, Naotaka ; Cornish, Virginia W. ; Margadant, Felix M. ; Sheetz, Michael. / Cooperative vinculin binding to talin mapped by time-resolved super resolution microscopy. In: Nano Letters. 2016 ; Vol. 16, No. 7. pp. 4062-4068.
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