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 Sheets

doi:10.1117/12.2571138

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 Sheets

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

2 Scopus citations

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.

Original language	English (US)
Title of host publication	Ultrafast Nonlinear Imaging and Spectroscopy VIII
Editors	Zhiwen Liu, Demetri Psaltis, Kebin Shi
Publisher	SPIE
ISBN (Electronic)	9781510638006
DOIs	https://doi.org/10.1117/12.2571138
State	Published - 2020
Event	Ultrafast Nonlinear Imaging and Spectroscopy VIII 2020 - Virtual, Online, United States Duration: Aug 24 2020 → Sep 4 2020

Publication series

Name	Proceedings of SPIE - The International Society for Optical Engineering
Volume	11497
ISSN (Print)	0277-786X
ISSN (Electronic)	1996-756X

Conference

Conference	Ultrafast Nonlinear Imaging and Spectroscopy VIII 2020
Country/Territory	United States
City	Virtual, Online
Period	8/24/20 → 9/4/20

Bibliographical note

Funding Information:
We would like to thank Robert Miller and Christin Libal for their technical help and useful discussion during the course of this project. E.D.S. and A.A.H. acknowledge the financial support provided by the University of Minnesota Grant-in-Aid, a Chancellor’s Small Grant, the Department of Chemistry and Biochemistry, the Swenson College of Science and Engineering, University of Minnesota Duluth. A.J.B. acknowledges the financial support of the Netherlands Organization for Scientific Research Vidi grant. C.P.A. was supported by teaching fellowships from the Department of Chemistry and Biochemistry, University of Minnesota Duluth. T.M.K. acknowledges the support of Mylan Radulovich Graduate Fellowship as well as the teaching fellowship from the Department of Physics and Astronomy, University of Minnesota Duluth. We further acknowledge the support from the Minnesota Supercomputing Institute (MSI) at the University of Minnesota.

Keywords

FRET
Fluorescence correlation spectroscopy
Time-resolved fluorescence
mCerulean3
mCitrine
mTurquoise2.1

Publisher link

10.1117/12.2571138

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Aplin, C. P., Kay, T. M., Beenken, J., Nwachuku, C., Tetteh-Jada, E., Heikal, A. A., Boersma, A. J., & Sheets, E. (2020). Comparative studies of the fluorescence spectroscopy and dynamics of mCerulean3 and mTurquoise2.1 as donors in FRET pairing with mCitrine. In Z. Liu, D. Psaltis, & K. Shi (Eds.), Ultrafast Nonlinear Imaging and Spectroscopy VIII Article 114970T (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 11497). SPIE. https://doi.org/10.1117/12.2571138

Comparative studies of the fluorescence spectroscopy and dynamics of mCerulean3 and mTurquoise2.1 as donors in FRET pairing with mCitrine. / Aplin, Cody P.; Kay, Taryn M.; Beenken, Julie et al.
Ultrafast Nonlinear Imaging and Spectroscopy VIII. ed. / Zhiwen Liu; Demetri Psaltis; Kebin Shi. SPIE, 2020. 114970T (Proceedings of SPIE - The International Society for Optical Engineering; Vol. 11497).

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Aplin, CP, Kay, TM, Beenken, J, Nwachuku, C, Tetteh-Jada, E, Heikal, AA, Boersma, AJ & Sheets, E 2020, Comparative studies of the fluorescence spectroscopy and dynamics of mCerulean3 and mTurquoise2.1 as donors in FRET pairing with mCitrine. in Z Liu, D Psaltis & K Shi (eds), Ultrafast Nonlinear Imaging and Spectroscopy VIII., 114970T, Proceedings of SPIE - The International Society for Optical Engineering, vol. 11497, SPIE, Ultrafast Nonlinear Imaging and Spectroscopy VIII 2020, Virtual, Online, United States, 8/24/20. https://doi.org/10.1117/12.2571138

Aplin CP, Kay TM, Beenken J, Nwachuku C, Tetteh-Jada E, Heikal AA et al. Comparative studies of the fluorescence spectroscopy and dynamics of mCerulean3 and mTurquoise2.1 as donors in FRET pairing with mCitrine. In Liu Z, Psaltis D, Shi K, editors, Ultrafast Nonlinear Imaging and Spectroscopy VIII. SPIE. 2020. 114970T. (Proceedings of SPIE - The International Society for Optical Engineering). doi: 10.1117/12.2571138

Aplin, Cody P. ; Kay, Taryn M. ; Beenken, Julie et al. / Comparative studies of the fluorescence spectroscopy and dynamics of mCerulean3 and mTurquoise2.1 as donors in FRET pairing with mCitrine. Ultrafast Nonlinear Imaging and Spectroscopy VIII. editor / Zhiwen Liu ; Demetri Psaltis ; Kebin Shi. SPIE, 2020. (Proceedings of SPIE - The International Society for Optical Engineering).

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

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 F{\"o}rster 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.",

keywords = "FRET, Fluorescence correlation spectroscopy, Time-resolved fluorescence, mCerulean3, mCitrine, mTurquoise2.1",

author = "Aplin, {Cody P.} and Kay, {Taryn M.} and Julie Beenken and Chioma Nwachuku and Emmanuel Tetteh-Jada and Heikal, {Ahmed A.} and Boersma, {Arnold J.} and Erin Sheets",

note = "Funding Information: We would like to thank Robert Miller and Christin Libal for their technical help and useful discussion during the course of this project. E.D.S. and A.A.H. acknowledge the financial support provided by the University of Minnesota Grant-in-Aid, a Chancellor{\textquoteright}s Small Grant, the Department of Chemistry and Biochemistry, the Swenson College of Science and Engineering, University of Minnesota Duluth. A.J.B. acknowledges the financial support of the Netherlands Organization for Scientific Research Vidi grant. C.P.A. was supported by teaching fellowships from the Department of Chemistry and Biochemistry, University of Minnesota Duluth. T.M.K. acknowledges the support of Mylan Radulovich Graduate Fellowship as well as the teaching fellowship from the Department of Physics and Astronomy, University of Minnesota Duluth. We further acknowledge the support from the Minnesota Supercomputing Institute (MSI) at the University of Minnesota.; Ultrafast Nonlinear Imaging and Spectroscopy VIII 2020 ; Conference date: 24-08-2020 Through 04-09-2020",

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doi = "10.1117/12.2571138",

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T1 - Comparative studies of the fluorescence spectroscopy and dynamics of mCerulean3 and mTurquoise2.1 as donors in FRET pairing with mCitrine

AU - Aplin, Cody P.

AU - Kay, Taryn M.

AU - Beenken, Julie

AU - Nwachuku, Chioma

AU - Tetteh-Jada, Emmanuel

AU - Heikal, Ahmed A.

AU - Boersma, Arnold J.

AU - Sheets, Erin

N1 - Funding Information: We would like to thank Robert Miller and Christin Libal for their technical help and useful discussion during the course of this project. E.D.S. and A.A.H. acknowledge the financial support provided by the University of Minnesota Grant-in-Aid, a Chancellor’s Small Grant, the Department of Chemistry and Biochemistry, the Swenson College of Science and Engineering, University of Minnesota Duluth. A.J.B. acknowledges the financial support of the Netherlands Organization for Scientific Research Vidi grant. C.P.A. was supported by teaching fellowships from the Department of Chemistry and Biochemistry, University of Minnesota Duluth. T.M.K. acknowledges the support of Mylan Radulovich Graduate Fellowship as well as the teaching fellowship from the Department of Physics and Astronomy, University of Minnesota Duluth. We further acknowledge the support from the Minnesota Supercomputing Institute (MSI) at the University of Minnesota.

PY - 2020

Y1 - 2020

N2 - 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.

AB - 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.

KW - FRET

KW - Fluorescence correlation spectroscopy

KW - Time-resolved fluorescence

KW - mCerulean3

KW - mCitrine

KW - mTurquoise2.1

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DO - 10.1117/12.2571138

M3 - Conference contribution

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T3 - Proceedings of SPIE - The International Society for Optical Engineering

BT - Ultrafast Nonlinear Imaging and Spectroscopy VIII

A2 - Liu, Zhiwen

A2 - Psaltis, Demetri

A2 - Shi, Kebin

PB - SPIE

T2 - Ultrafast Nonlinear Imaging and Spectroscopy VIII 2020

Y2 - 24 August 2020 through 4 September 2020

ER -

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

Abstract

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Bibliographical note

Keywords

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