A Combination of Library Screening and Rational Mutagenesis Expands the Available Color Palette of the Smallest Fluorogen-Activating Protein Tag nanoFAST

NanoFAST is the smallest fluorogen-activating protein, consisting of only 98 amino acids, used as a genetically encoded fluorescent tag. Previously, only a single fluorogen with an orange color was revealed for this protein. In the present paper, using rational mutagenesis and in vitro screening of...

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Published in:International journal of molecular sciences 2024-03, Vol.25 (5), p.3054
Main Authors: Baleeva, Nadezhda S, Bogdanova, Yulia A, Goncharuk, Marina V, Sokolov, Anatolii I, Myasnyanko, Ivan N, Kublitski, Vadim S, Smirnov, Alexander Yu, Gilvanov, Aidar R, Goncharuk, Sergey A, Mineev, Konstantin S, Baranov, Mikhail S
Format: Article
Language:English
Subjects:
Amino acids
Electronics industry
FAST
fluorescence
Fluorescent Dyes - chemistry
fluorogen
fluorogen-activating proteins
Libraries
Ligands
Light emitting diodes
Microscopy
Mutagenesis
nanoFAST
Proteins
Visualization
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Summary:NanoFAST is the smallest fluorogen-activating protein, consisting of only 98 amino acids, used as a genetically encoded fluorescent tag. Previously, only a single fluorogen with an orange color was revealed for this protein. In the present paper, using rational mutagenesis and in vitro screening of fluorogens libraries, we expanded the color palette of this tag. We discovered that E46Q is one of the key substitutions enabling the range of possible fluorogens to be expanded. The introduction of this and several other substitutions has made it possible to use not only orange but also red and green fluorogens with the modified protein.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms25053054