Single molecule fluctuation dynamics reveal protein size, structure and affinities
Principal investigator : Jerome Wenger
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Principal investigator : Jerome Wenger
Fluorescence Correlation Spectroscopy uses a confocal microscope to collect the light from luminescent molecules moving across a femtoliter observation volume (one femtoliter volume is one micron cube). The fluctuating number of fluorophores in this observation volume gives rise to intensity fluctuations which provide information about diffusion, molecular concentration and associations. FRET uses donor-acceptor fluorescence energy transfer as nanoscopic ruler to monitor conformation changes and association dynamics.
We apply FCS and FRET to monitor the interaction dynamics of various proteins, including :
[4] S. B. Moparthi, U. Carlsson, R. Vincentelli, B.-H. Jonsson, P. Hammarström, J. Wenger, Differential conformational modulations of MreB folding upon interactions with GroEL/ES and TRiC chaperonin components, Sci. Rep. 6, 28386 (2016).
[3] S.B. Moparthi, G. Thieulin-Pardo, J. de Torres, P. Ghenuche, B. Gontero, J. Wenger, FRET analysis of CP12 structural interplay by GAPDH and PRK, Biochem. Biophys. Res. Comm. 458, 488-493 (2015).
[2] S.B. Moparthi, G. Thieulin-Pardo, P. Mansuelle, H. Rigneault, B. Gontero, J. Wenger, Conformational modulation and hydrodynamic radii of CP12 protein and its complexes probed by fluorescence correlation spectroscopy, FEBS Journal 281, 3206-3217 (2014)
[1] C. Favard, J. Wenger, P.-F. Lenne, H. Rigneault, FCS diffusion laws on two-phases lipid membranes : experimental and Monte-carlo simulations determination of domain mean size, Biophys. J. 100, 1242-1251 (2011).