Green fluorescent protein with anionic tryptophan-based chromophore and long fluorescence lifetime.
Biophys J
; 109(2): 380-9, 2015 Jul 21.
Article
em En
| MEDLINE
| ID: mdl-26200874
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Proteínas de Fluorescência Verde
Limite:
Animals
/
Humans
Idioma:
En
Revista:
Biophys J
Ano de publicação:
2015
Tipo de documento:
Article
País de afiliação:
Federação Russa