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Journal Article

Dynamic tuning of FRET in a green fluorescent protein biosensor

MPS-Authors
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Thestrup,  Thomas
Research Group: Tools for Bio-Imaging / Griesbeck, MPI of Neurobiology, Max Planck Society;

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Griesbeck,  Oliver
Research Group: Tools for Bio-Imaging / Griesbeck, MPI of Neurobiology, Max Planck Society;

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eaaw4988.full.pdf
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Citation

Trigo-Mourino, P., Thestrup, T., Griesbeck, O., Griesinger, C., & Becker, S. (2019). Dynamic tuning of FRET in a green fluorescent protein biosensor. Science Advances, 5(8): eaaw4988. doi:10.1126/sciadv.aaw4988.


Cite as: http://hdl.handle.net/21.11116/0000-0005-ACCA-6
Abstract
Forster resonance energy transfer (FRET) between mutants of green fluorescent protein is widely used to monitor protein-protein interactions and as a readout mode in fluorescent biosensors. Despite the fundamental importance of distance and molecular angles of fluorophores to each other, structural details on fluorescent protein FRET have been missing. Here, we report the high-resolution x-ray structure of the fluorescent proteins mCerulean3 and cpVenus within the biosensor Twitch-2B, as they undergo FRET and characterize the dynamics of this biosensor with B-0(2)-dependent paramagnetic nuclear magnetic resonance at 900 MHz and 1.1 GHz. These structural data provide the unprecedented opportunity to calculate FRET from the x-ray structure and to compare it to experimental data in solution. We find that interdomain dynamics limits the FRET effect and show that a rigidification of the sensor further enhances FRET.