Comparative studies of the fluorescence spectroscopy and dynamics of mCerulean3 and mTurquoise2.1 as donors in FRET pairing with mCitrine

Cody P. Aplin; Taryn M. Kay; Julie Beenken; Chioma Nwachuku; Emmanuel Tetteh-Jada; Ahmed A. Heikal; Arnold J. Boersma; Erin D. Sheets

doi:10.1117/12.2571138

20 August 2020 Comparative studies of the fluorescence spectroscopy and dynamics of mCerulean3 and mTurquoise2.1 as donors in FRET pairing with mCitrine

Cody P. Aplin, Taryn M. Kay, Julie Beenken, Chioma Nwachuku, Emmanuel Tetteh-Jada, Ahmed A. Heikal, Arnold J. Boersma, Erin D. Sheets

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Proceedings Volume 11497, Ultrafast Nonlinear Imaging and Spectroscopy VIII; 114970T (2020) https://doi.org/10.1117/12.2571138
Event: SPIE Optical Engineering + Applications, 2020, Online Only

Abstract

We investigated the donor effects (mCerulean3 versus mTurquoise2.1) on the spectroscopy and dynamics of mCerulean3-linker-mCitrine constructs using integrated fluorescence spectroscopy methods. Here, mCerulean3 (a cyan fluorescent protein) and mCitrine (a yellow fluorescent protein) act as Förster resonance energy transfer (FRET) pair, separated by flexible linker region. We hypothesize that the construct with mTurquoise2.1 would have many advantages as a donor, which include a higher FRET efficiency as compared with the mCerulean3 due to the enhanced spectral overlap with mCitrine. To test this hypothesis, we used steady-state spectroscopy, time-resolved fluorescence, and fluorescence correlation spectroscopy of both mCerulean3-linker-mCitrine and mTurquoise2.1-linker-mCitrine to investigate the donor effect on the FRET efficiency and translational diffusion as a means for developing a rational design for hetero-FRET constructs for environmental sensing.

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Cody P. Aplin, Taryn M. Kay, Julie Beenken, Chioma Nwachuku, Emmanuel Tetteh-Jada, Ahmed A. Heikal, Arnold J. Boersma, and Erin D. Sheets "Comparative studies of the fluorescence spectroscopy and dynamics of mCerulean3 and mTurquoise2.1 as donors in FRET pairing with mCitrine", Proc. SPIE 11497, Ultrafast Nonlinear Imaging and Spectroscopy VIII, 114970T (20 August 2020); https://doi.org/10.1117/12.2571138

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