Green Fluorescent Protein with Anionic Tryptophan-Based Chromophore and Long Fluorescence Lifetime

Karen S. Sarkisyan; Alexander S. Goryashchenko; Peter V. Lidsky; Dmitry A. Gorbachev; Nina G. Bozhanova; Andrey Yu. Gorokhovatsky; Alina R. Pereverzeva; Alina P. Ryumina; Victoria V. Zherdeva; Alexander P. Savitsky; Kyril M. Solntsev; Andreas S. Bommarius; George V. Sharonov; Jake R. Lindquist; Mikhail Drobizhev; Thomas E. Hughes; Aleksander Rebane; Konstantin A. Lukyanov; Alexander S. Mishin

doi:10.1016/j.bpj.2015.06.018

Green Fluorescent Protein with Anionic Tryptophan-Based Chromophore and Long Fluorescence Lifetime

Karen S. Sarkisyan, Alexander S. Goryashchenko, Peter V. Lidsky, Dmitry A. Gorbachev, Nina G. Bozhanova, Andrey Yu. Gorokhovatsky, Alina R. Pereverzeva, Alina P. Ryumina, Victoria V. Zherdeva, Alexander P. Savitsky, Kyril M. Solntsev, Andreas S. Bommarius, George V. Sharonov, Jake R. Lindquist, Mikhail Drobizhev, Thomas E. Hughes, Aleksander Rebane, Konstantin A. Lukyanov, Alexander S. Mishin^*

^*Corresponding author for this work

Research output: Journal Publications and Reviews › RGC 21 - Publication in refereed journal › peer-review

55 Citations (Scopus)

Abstract

Spectral diversity of fluorescent proteins, crucial for multiparameter imaging, is based mainly on chemical diversity of their chromophores. Recently we have reported, to our knowledge, a new green fluorescent protein WasCFP - the first fluorescent protein with a tryptophan-based chromophore in the anionic state. However, only a small portion of WasCFP molecules exists in the anionic state at physiological conditions. In this study we report on an improved variant of WasCFP, named NowGFP, with the anionic form dominating at 37°C and neutral pH. It is 30% brighter than enhanced green fluorescent protein (EGFP) and exhibits a fluorescence lifetime of 5.1 ns. We demonstrated that signals of NowGFP and EGFP can be clearly distinguished by fluorescence lifetime in various models, including mammalian cells, mouse tumor xenograft, and Drosophila larvae. NowGFP thus provides an additional channel for multiparameter fluorescence lifetime imaging microscopy of green fluorescent proteins. © 2015 Biophysical Society.

Original language	English
Pages (from-to)	380-389
Journal	Biophysical Journal
Volume	109
Issue number	2
DOIs	https://doi.org/10.1016/j.bpj.2015.06.018
Publication status	Published - 22 Jul 2015
Externally published	Yes

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Sarkisyan, K. S., Goryashchenko, A. S., Lidsky, P. V., Gorbachev, D. A., Bozhanova, N. G., Gorokhovatsky, A. Y., Pereverzeva, A. R., Ryumina, A. P., Zherdeva, V. V., Savitsky, A. P., Solntsev, K. M., Bommarius, A. S., Sharonov, G. V., Lindquist, J. R., Drobizhev, M., Hughes, T. E., Rebane, A., Lukyanov, K. A., & Mishin, A. S. (2015). Green Fluorescent Protein with Anionic Tryptophan-Based Chromophore and Long Fluorescence Lifetime. Biophysical Journal, 109(2), 380-389. https://doi.org/10.1016/j.bpj.2015.06.018

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title = "Green Fluorescent Protein with Anionic Tryptophan-Based Chromophore and Long Fluorescence Lifetime",

abstract = "Spectral diversity of fluorescent proteins, crucial for multiparameter imaging, is based mainly on chemical diversity of their chromophores. Recently we have reported, to our knowledge, a new green fluorescent protein WasCFP - the first fluorescent protein with a tryptophan-based chromophore in the anionic state. However, only a small portion of WasCFP molecules exists in the anionic state at physiological conditions. In this study we report on an improved variant of WasCFP, named NowGFP, with the anionic form dominating at 37°C and neutral pH. It is 30% brighter than enhanced green fluorescent protein (EGFP) and exhibits a fluorescence lifetime of 5.1 ns. We demonstrated that signals of NowGFP and EGFP can be clearly distinguished by fluorescence lifetime in various models, including mammalian cells, mouse tumor xenograft, and Drosophila larvae. NowGFP thus provides an additional channel for multiparameter fluorescence lifetime imaging microscopy of green fluorescent proteins. {\textcopyright} 2015 Biophysical Society.",

author = "Sarkisyan, {Karen S.} and Goryashchenko, {Alexander S.} and Lidsky, {Peter V.} and Gorbachev, {Dmitry A.} and Bozhanova, {Nina G.} and Gorokhovatsky, {Andrey Yu.} and Pereverzeva, {Alina R.} and Ryumina, {Alina P.} and Zherdeva, {Victoria V.} and Savitsky, {Alexander P.} and Solntsev, {Kyril M.} and Bommarius, {Andreas S.} and Sharonov, {George V.} and Lindquist, {Jake R.} and Mikhail Drobizhev and Hughes, {Thomas E.} and Aleksander Rebane and Lukyanov, {Konstantin A.} and Mishin, {Alexander S.}",

note = "Publication details (e.g. title, author(s), publication statuses and dates) are captured on an “AS IS” and “AS AVAILABLE” basis at the time of record harvesting from the data source. Suggestions for further amendments or supplementary information can be sent to [email protected].",

year = "2015",

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doi = "10.1016/j.bpj.2015.06.018",

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Sarkisyan, KS, Goryashchenko, AS, Lidsky, PV, Gorbachev, DA, Bozhanova, NG, Gorokhovatsky, AY, Pereverzeva, AR, Ryumina, AP, Zherdeva, VV, Savitsky, AP, Solntsev, KM, Bommarius, AS, Sharonov, GV, Lindquist, JR, Drobizhev, M, Hughes, TE, Rebane, A, Lukyanov, KA & Mishin, AS 2015, 'Green Fluorescent Protein with Anionic Tryptophan-Based Chromophore and Long Fluorescence Lifetime', Biophysical Journal, vol. 109, no. 2, pp. 380-389. https://doi.org/10.1016/j.bpj.2015.06.018

TY - JOUR

T1 - Green Fluorescent Protein with Anionic Tryptophan-Based Chromophore and Long Fluorescence Lifetime

AU - Sarkisyan, Karen S.

AU - Goryashchenko, Alexander S.

AU - Lidsky, Peter V.

AU - Gorbachev, Dmitry A.

AU - Bozhanova, Nina G.

AU - Gorokhovatsky, Andrey Yu.

AU - Pereverzeva, Alina R.

AU - Ryumina, Alina P.

AU - Zherdeva, Victoria V.

AU - Savitsky, Alexander P.

AU - Solntsev, Kyril M.

AU - Bommarius, Andreas S.

AU - Sharonov, George V.

AU - Lindquist, Jake R.

AU - Drobizhev, Mikhail

AU - Hughes, Thomas E.

AU - Rebane, Aleksander

AU - Lukyanov, Konstantin A.

AU - Mishin, Alexander S.

N1 - Publication details (e.g. title, author(s), publication statuses and dates) are captured on an “AS IS” and “AS AVAILABLE” basis at the time of record harvesting from the data source. Suggestions for further amendments or supplementary information can be sent to [email protected].

PY - 2015/7/22

Y1 - 2015/7/22

N2 - Spectral diversity of fluorescent proteins, crucial for multiparameter imaging, is based mainly on chemical diversity of their chromophores. Recently we have reported, to our knowledge, a new green fluorescent protein WasCFP - the first fluorescent protein with a tryptophan-based chromophore in the anionic state. However, only a small portion of WasCFP molecules exists in the anionic state at physiological conditions. In this study we report on an improved variant of WasCFP, named NowGFP, with the anionic form dominating at 37°C and neutral pH. It is 30% brighter than enhanced green fluorescent protein (EGFP) and exhibits a fluorescence lifetime of 5.1 ns. We demonstrated that signals of NowGFP and EGFP can be clearly distinguished by fluorescence lifetime in various models, including mammalian cells, mouse tumor xenograft, and Drosophila larvae. NowGFP thus provides an additional channel for multiparameter fluorescence lifetime imaging microscopy of green fluorescent proteins. © 2015 Biophysical Society.

AB - Spectral diversity of fluorescent proteins, crucial for multiparameter imaging, is based mainly on chemical diversity of their chromophores. Recently we have reported, to our knowledge, a new green fluorescent protein WasCFP - the first fluorescent protein with a tryptophan-based chromophore in the anionic state. However, only a small portion of WasCFP molecules exists in the anionic state at physiological conditions. In this study we report on an improved variant of WasCFP, named NowGFP, with the anionic form dominating at 37°C and neutral pH. It is 30% brighter than enhanced green fluorescent protein (EGFP) and exhibits a fluorescence lifetime of 5.1 ns. We demonstrated that signals of NowGFP and EGFP can be clearly distinguished by fluorescence lifetime in various models, including mammalian cells, mouse tumor xenograft, and Drosophila larvae. NowGFP thus provides an additional channel for multiparameter fluorescence lifetime imaging microscopy of green fluorescent proteins. © 2015 Biophysical Society.

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DO - 10.1016/j.bpj.2015.06.018

M3 - RGC 21 - Publication in refereed journal

C2 - 26200874

SN - 0006-3495

VL - 109

SP - 380

EP - 389

JO - Biophysical Journal

JF - Biophysical Journal

IS - 2

ER -

Green Fluorescent Protein with Anionic Tryptophan-Based Chromophore and Long Fluorescence Lifetime

Abstract

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