Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy

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 dynamic...

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Autores principales:	Corbat, A.A., Schuermann, K.C., Liguzinski, P., Radon, Y., Bastiaens, P.I.H., Verveer, P.J., Grecco, H.E.
Formato:	JOUR
Materias:	Anisotropy FRET biosensor Apoptotic network Caspase activity Co-monitoring Imaging Polarization microscopy caspase 3 caspase 8 caspase 9 effector caspase anisotropy anisotropy forster resonant energy transfer based biosensor apoptosis Article controlled study correlation analysis enzyme activation enzyme activity fluorescence anisotropy microscopy human human cell microscopy molecular model priority journal simulation fluorescence microscopy fluorescence polarization fluorescence resonance energy transfer genetic procedures HeLa cell line metabolism procedures signal transduction Apoptosis Biosensing Techniques Caspases, Effector Enzyme Activation Fluorescence Polarization Fluorescence Resonance Energy Transfer HeLa Cells Humans Microscopy, Fluorescence Signal Transduction
Acceso en línea:	http://hdl.handle.net/20.500.12110/paper_22132317_v19_n_p210_Corbat
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	todo:paper_22132317_v19_n_p210_Corbat
record_format	dspace
spelling	todo:paper_22132317_v19_n_p210_Corbat2023-10-03T16:40:35Z Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy Corbat, A.A. Schuermann, K.C. Liguzinski, P. Radon, Y. Bastiaens, P.I.H. Verveer, P.J. Grecco, H.E. Anisotropy FRET biosensor Apoptotic network Caspase activity Co-monitoring Imaging Polarization microscopy caspase 3 caspase 8 caspase 9 effector caspase anisotropy anisotropy forster resonant energy transfer based biosensor apoptosis Article controlled study correlation analysis enzyme activation enzyme activity fluorescence anisotropy microscopy human human cell microscopy molecular model priority journal simulation apoptosis fluorescence microscopy fluorescence polarization fluorescence resonance energy transfer genetic procedures HeLa cell line metabolism procedures signal transduction Apoptosis Biosensing Techniques Caspases, Effector Enzyme Activation Fluorescence Polarization Fluorescence Resonance Energy Transfer HeLa Cells Humans Microscopy, Fluorescence Signal Transduction 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. © 2018 JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_22132317_v19_n_p210_Corbat
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	Anisotropy FRET biosensor Apoptotic network Caspase activity Co-monitoring Imaging Polarization microscopy caspase 3 caspase 8 caspase 9 effector caspase anisotropy anisotropy forster resonant energy transfer based biosensor apoptosis Article controlled study correlation analysis enzyme activation enzyme activity fluorescence anisotropy microscopy human human cell microscopy molecular model priority journal simulation apoptosis fluorescence microscopy fluorescence polarization fluorescence resonance energy transfer genetic procedures HeLa cell line metabolism procedures signal transduction Apoptosis Biosensing Techniques Caspases, Effector Enzyme Activation Fluorescence Polarization Fluorescence Resonance Energy Transfer HeLa Cells Humans Microscopy, Fluorescence Signal Transduction
spellingShingle	Anisotropy FRET biosensor Apoptotic network Caspase activity Co-monitoring Imaging Polarization microscopy caspase 3 caspase 8 caspase 9 effector caspase anisotropy anisotropy forster resonant energy transfer based biosensor apoptosis Article controlled study correlation analysis enzyme activation enzyme activity fluorescence anisotropy microscopy human human cell microscopy molecular model priority journal simulation apoptosis fluorescence microscopy fluorescence polarization fluorescence resonance energy transfer genetic procedures HeLa cell line metabolism procedures signal transduction Apoptosis Biosensing Techniques Caspases, Effector Enzyme Activation Fluorescence Polarization Fluorescence Resonance Energy Transfer HeLa Cells Humans Microscopy, Fluorescence Signal Transduction Corbat, A.A. Schuermann, K.C. Liguzinski, P. Radon, Y. Bastiaens, P.I.H. Verveer, P.J. Grecco, H.E. Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy
topic_facet	Anisotropy FRET biosensor Apoptotic network Caspase activity Co-monitoring Imaging Polarization microscopy caspase 3 caspase 8 caspase 9 effector caspase anisotropy anisotropy forster resonant energy transfer based biosensor apoptosis Article controlled study correlation analysis enzyme activation enzyme activity fluorescence anisotropy microscopy human human cell microscopy molecular model priority journal simulation apoptosis fluorescence microscopy fluorescence polarization fluorescence resonance energy transfer genetic procedures HeLa cell line metabolism procedures signal transduction Apoptosis Biosensing Techniques Caspases, Effector Enzyme Activation Fluorescence Polarization Fluorescence Resonance Energy Transfer HeLa Cells Humans Microscopy, Fluorescence Signal Transduction
description	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. © 2018
format	JOUR
author	Corbat, A.A. Schuermann, K.C. Liguzinski, P. Radon, Y. Bastiaens, P.I.H. Verveer, P.J. Grecco, H.E.
author_facet	Corbat, A.A. Schuermann, K.C. Liguzinski, P. Radon, Y. Bastiaens, P.I.H. Verveer, P.J. Grecco, H.E.
author_sort	Corbat, A.A.
title	Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy
title_short	Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy
title_full	Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy
title_fullStr	Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy
title_full_unstemmed	Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy
title_sort	co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy
url	http://hdl.handle.net/20.500.12110/paper_22132317_v19_n_p210_Corbat
work_keys_str_mv	AT corbataa coimagingextrinsicintrinsicandeffectorcaspaseactivitybyfluorescenceanisotropymicroscopy AT schuermannkc coimagingextrinsicintrinsicandeffectorcaspaseactivitybyfluorescenceanisotropymicroscopy AT liguzinskip coimagingextrinsicintrinsicandeffectorcaspaseactivitybyfluorescenceanisotropymicroscopy AT radony coimagingextrinsicintrinsicandeffectorcaspaseactivitybyfluorescenceanisotropymicroscopy AT bastiaenspih coimagingextrinsicintrinsicandeffectorcaspaseactivitybyfluorescenceanisotropymicroscopy AT verveerpj coimagingextrinsicintrinsicandeffectorcaspaseactivitybyfluorescenceanisotropymicroscopy AT greccohe coimagingextrinsicintrinsicandeffectorcaspaseactivitybyfluorescenceanisotropymicroscopy
_version_	1782027353187155968

Co-imaging extrinsic, intrinsic and effector caspase activity by fluorescence anisotropy microscopy

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