Imperial College London

ProfessorPaulFrench

Faculty of Natural SciencesDepartment of Physics

Professor of Physics and Vice Dean (Research) - FoNS
 
 
 
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Contact

 

+44 (0)20 7594 7706paul.french Website

 
 
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Assistant

 

Ms Judith Baylis +44 (0)20 7594 7713

 
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Location

 

609Blackett LaboratorySouth Kensington Campus

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Summary

 

Publications

Citation

BibTex format

@article{Margineanu:2016:10.1038/srep28186,
author = {Margineanu, A and Chan, JJ and Kelly, DJ and Warren, S and Flatters, D and Kumar, S and Katan, M and Dunsby, C and French, PMW},
doi = {10.1038/srep28186},
journal = {Scientific Reports},
title = {Screening for protein-protein interactions using Förster resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM)},
url = {http://dx.doi.org/10.1038/srep28186},
volume = {6},
year = {2016}
}

RIS format (EndNote, RefMan)

TY  - JOUR
AB - We present a high content multiwell plate cell-based assay approach to quantify protein interactions directly in cells using Förster resonance energy transfer (FRET) read out by automated fluorescence lifetime imaging (FLIM). Automated FLIM is implemented using wide-field time-gated detection, typically requiring only 10 s per field of view (FOV). Averaging over biological, thermal and shot noise with 100’s to 1000’s of FOV enables unbiased quantitative analysis with high statistical power. Plotting average donor lifetime vs. acceptor/donor intensity ratio clearly identifies protein interactions and fitting to double exponential donor decay models provides estimates of interacting population fractions that, with calibrated donor and acceptor fluorescence intensities, can yield dissociation constants. We demonstrate the application to identify binding partners of MST1 kinase and estimate interaction strength among the members of the RASSF protein family, which have important roles in apoptosis via the Hippo signalling pathway. KD values broadly agree with published biochemical measurements.
AU - Margineanu,A
AU - Chan,JJ
AU - Kelly,DJ
AU - Warren,S
AU - Flatters,D
AU - Kumar,S
AU - Katan,M
AU - Dunsby,C
AU - French,PMW
DO - 10.1038/srep28186
PY - 2016///
SN - 2045-2322
TI - Screening for protein-protein interactions using Förster resonance energy transfer (FRET) and fluorescence lifetime imaging microscopy (FLIM)
T2 - Scientific Reports
UR - http://dx.doi.org/10.1038/srep28186
UR - http://hdl.handle.net/10044/1/33008
VL - 6
ER -