Quantifying intracellular protein binding thermodynamics during mechanotransduction based on FRET spectroscopy.

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Serval ID
serval:BIB_9FFA0A07FCDE
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Quantifying intracellular protein binding thermodynamics during mechanotransduction based on FRET spectroscopy.
Journal
Methods
Author(s)
Abdul Rahim N.A., Pelet S., Mofrad M.R., So P.T., Kamm R.D.
ISSN
1095-9130 (Electronic)
ISSN-L
1046-2023
Publication state
Published
Issued date
2014
Peer-reviewed
Oui
Volume
66
Number
2
Pages
208-221
Language
english
Notes
Publication types: Journal Article
Abstract
Mechanical force modulates myriad cellular functions including migration, alignment, proliferation, and gene transcription. Mechanotransduction, the transmission of mechanical forces and its translation into biochemical signals, may be mediated by force induced protein conformation changes, subsequently modulating protein signaling. For the paxillin and focal adhesion kinase interaction, we demonstrate that force-induced changes in protein complex conformation, dissociation constant, and binding Gibbs free energy can be quantified by lifetime-resolved fluorescence energy transfer microscopy combined with intensity imaging calibrated by fluorescence correlation spectroscopy. Comparison with in vitro data shows that this interaction is allosteric in vivo. Further, spatially resolved imaging and inhibitor assays show that this protein interaction and its mechano-sensitivity are equal in the cytosol and in the focal adhesions complexes indicating that the mechano-sensitivity of this interaction must be mediated by soluble factors but not based on protein tyrosine phosphorylation.
Pubmed
Web of science
Open Access
Yes
Create date
08/01/2014 17:43
Last modification date
20/08/2019 15:06
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