NanoFAST: structure-based design of a small fluorogen-activating protein with only 98 amino acids.
Konstantin S. Mineev,Konstantin S. Mineev,Sergey A. Goncharuk,Sergey A. Goncharuk,Marina V. Goncharuk,Natalia V. Povarova,Anatolii I. Sokolov,Nadezhda S. Baleeva,Alexander Yu. Smirnov,Ivan N. Myasnyanko,Dmitry A. Ruchkin,Sergey Bukhdruker,Alina Remeeva,Alexey Mishin,Valentin Borshchevskiy,Valentin Borshchevskiy,Valentin Gordeliy,Alexander S. Arseniev,Dmitriy A. Gorbachev,Alexey S. Gavrikov,Alexander S. Mishin,Mikhail S. Baranov,Mikhail S. Baranov +22 more
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TLDR
The shortened FAST is designed, which is composed of only 98 amino acids, the shortest genetically encoded tag among all known fluorescent and fluorogen-activating proteins, by truncating 26 N-terminal residues.Abstract:
One of the essential characteristics of any tag used in bioscience and medical applications is its size. The larger the label, the more it may affect the studied object, and the more it may distort its behavior. In this paper, using NMR spectroscopy and X-ray crystallography, we have studied the structure of fluorogen-activating protein FAST both in the apo form and in complex with the fluorogen. We showed that significant change in the protein occurs upon interaction with the ligand. While the protein is completely ordered in the complex, its apo form is characterized by higher mobility and disordering of its N-terminus. We used structural information to design the shortened FAST (which we named nanoFAST) by truncating 26 N-terminal residues. Thus, we created the shortest genetically encoded tag among all known fluorescent and fluorogen-activating proteins, which is composed of only 98 amino acids.read more
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Engineering of a fluorescent chemogenetic reporter with tunable color for advanced live-cell imaging
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TL;DR: In this paper , the structure-based rational design of the enhanced FAST mutants, optimized for the N871b fluorogen, was described, and two mutants appeared brighter than the wild-type FAST, and these mutants provided up to 35% enhancement for several other fluorogens of similar structure, both in vitro and in vivo.
References
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Journal ArticleDOI
Improved chemical-genetic fluorescent markers for live cell microscopy
TL;DR: Rational design is used to modify the binding pocket of the protein and screen for improved fluorescence performances with four different fluorogens, resulting in improvements in both quantum yield and dissociation constant with nearly all fluorogens tested.
Journal ArticleDOI
A Far‐Red Emitting Fluorescent Chemogenetic Reporter for In Vivo Molecular Imaging
Chenge Li,Chenge Li,Alison G. Tebo,Marion Thauvin,Marion Thauvin,Marie-Aude Plamont,Michel Volovitch,Michel Volovitch,Xavier Morin,Sophie Vriz,Sophie Vriz,Arnaud Gautier,Arnaud Gautier +12 more
TL;DR: The generation of frFAST, a 14-kDa monomeric protein that forms a bright far-red fluorescent assembly with (4-hydroxy-3-methoxy-phenyl)allylidene rhodanine (HPAR-3OM), allowed the design of a far- red chemogenetic reporter of protein-protein interactions, demonstrating its great potential for theDesign of innovative far-Red emitting biosensors.
Journal ArticleDOI
A far‐red fluorescent chemogenetic reporter for in vivo molecular imaging
Chenge Li,Alison G. Tebo,Marion Thauvin,Marie-Aude Plamont,Michel Volovitch,Xavier Morin,Sophie Vriz,Arnaud Gautier +7 more
TL;DR: The frFAST (far-red fluorescence activating and absorption shifting tag) as mentioned in this paper is a 14-kDa monomeric protein that forms a bright far-red fluorescent assembly with (4-hydroxy-3-methoxy-phenyl)allylidene rhodanine (HPAR-3OM).
Journal ArticleDOI
Fluorophore tagged bio-molecules and their applications: A brief review
Ivy Sarkar,Ashok Kumar Mishra +1 more
TL;DR: This review demonstrates the applications of such conjugated fluorescent molecular probes in different domains of biological activities and brings out the advantages and disadvantages of this particular type of fluorophores with the insight to the future perspectives.
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Announcement of the BioSync web site.
TL;DR: This resource serves as a structural biologist’s guide to synchrotron facilities and is a part of the National Biomedical Computation Resource, an NIH National Center for Research Resources hosted at the San Diego Supercomputer Center.