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

Research output: Contribution to journalArticlepeer-review

36 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.

Original languageEnglish
Pages (from-to)380-389
Number of pages10
JournalBiophysical Journal
Volume109
Issue number2
DOIs
Publication statusPublished - 22 Jul 2015
Externally publishedYes

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