Abstract:
:In order to overcome intercellular variability and thereby effectively assess signal propagation in biological networks it is imperative to simultaneously quantify multiple biological observables in single living cells. While fluorescent biosensors have been the tool of choice to monitor the dynamics of protein interaction and enzymatic activity, co-measuring more than two of them has proven challenging. In this work, we designed three spectrally separated anisotropy-based Förster Resonant Energy Transfer (FRET) biosensors to overcome this difficulty. We demonstrate this principle by monitoring the activation of extrinsic, intrinsic and effector caspases upon apoptotic stimulus. Together with modelling and simulations we show that time of maximum activity for each caspase can be derived from the anisotropy of the corresponding biosensor. Such measurements correlate relative activation times and refine existing models of biological signalling networks, providing valuable insight into signal propagation.
journal_name
Redox Bioljournal_title
Redox biologyauthors
Corbat AA,Schuermann KC,Liguzinski P,Radon Y,Bastiaens PIH,Verveer PJ,Grecco HEdoi
10.1016/j.redox.2018.07.023subject
Has Abstractpub_date
2018-10-01 00:00:00pages
210-217issn
2213-2317pii
S2213-2317(18)30552-4journal_volume
19pub_type
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