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  Multiplexing molecular tension sensors reveals piconewton force gradient across talin-1

Ringer, P., Weissl, A., Cost, A.-L., Freikamp, A., Sabass, B., Mehlich, A., et al. (2017). Multiplexing molecular tension sensors reveals piconewton force gradient across talin-1. Nature methods, 14(11), 1090-1096. doi:10.1038/NMETH.4431.

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 Creators:
Ringer, Pia1, Author           
Weissl, Andreas2, Author
Cost, Anna-Lena1, Author           
Freikamp, Andrea1, Author           
Sabass, Benedikt2, Author
Mehlich, Alexander2, Author
Tramier, Marc2, Author
Rief, Matthias2, Author
Grashoff, Carsten1, Author           
Affiliations:
1Grashoff, Carsten / Molecular Mechanotransduction, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565150              
2external, ou_persistent22              

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Free keywords: FOCAL ADHESION DYNAMICS; FLUORESCENT PROTEIN; OPTICAL TWEEZERS; INTEGRIN; CELLS; TRANSDUCTION; BIOSENSORS; RESOLUTION; MECHANICS; RIGIDITYBiochemistry & Molecular Biology;
 Abstract: Forster resonance energy transfer (FRET)-based tension sensor modules (TSM s) are available for investigating how distinct proteins bear mechanical forces in cells. Yet, forces in the single piconewton (pN) regime remain difficult to resolve, and tools for multiplexed tension sensing are lacking. Here, we report the generation and calibration of a genetically encoded, FRET -based biosensor called FL-TSM, which is characterized by a near-digital force response and increased sensitivity at 3-5 pN. In addition, we present a method allowing the simultaneous evaluation of coexpressed tension sensor constructs using two-color fluorescence lifetime microscopy. Finally, we introduce a procedure to calculate the fraction of mechanically engaged molecules within cells. Application of these techniques to new talin biosensors reveals an intramolecular tension gradient across talin-1 that is established upon integrin-mediated cell adhesion. The tension gradient is actomyosin-and vinculin-dependent and sensitive to the rigidity of the extracellular environment.

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Language(s): eng - English
 Dates: 2017
 Publication Status: Published in print
 Pages: 13
 Publishing info: -
 Table of Contents: -
 Rev. Type: -
 Identifiers: ISI: 000414120400024
DOI: 10.1038/NMETH.4431
 Degree: -

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Title: Nature methods
  Other : Nature methods
Source Genre: Journal
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Publ. Info: New York, NY : Nature Pub. Group
Pages: - Volume / Issue: 14 (11) Sequence Number: - Start / End Page: 1090 - 1096 Identifier: ISSN: 1548-7091
CoNE: https://pure.mpg.de/cone/journals/resource/111088195279556