The role of the correlated motion(s) of the chromophore in photoswitching of green and red forms of the photoconvertible fluorescent protein mSAASoti

Abstract Wild-type SAASoti and its monomeric variant mSAASoti can undergo phototransformations, including reversible photoswitching of the green form to a nonfluorescent state and irreversible green-to-red photoconversion. In this study, we extend the photochemistry of mSAASoti variants to enable re...

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Main Authors: Alexandra V. Gavshina, Ilya D. Solovyev, Maria G. Khrenova, Konstantin M. Boyko, Larisa A. Varfolomeeva, Mikhail E. Minyaev, Vladimir O. Popov, Alexander P. Savitsky
Format: Article
Language:English
Published: Nature Portfolio 2024-04-01
Series:Scientific Reports
Subjects:
Online Access:https://doi.org/10.1038/s41598-024-59364-1
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author Alexandra V. Gavshina
Ilya D. Solovyev
Maria G. Khrenova
Konstantin M. Boyko
Larisa A. Varfolomeeva
Mikhail E. Minyaev
Vladimir O. Popov
Alexander P. Savitsky
author_facet Alexandra V. Gavshina
Ilya D. Solovyev
Maria G. Khrenova
Konstantin M. Boyko
Larisa A. Varfolomeeva
Mikhail E. Minyaev
Vladimir O. Popov
Alexander P. Savitsky
author_sort Alexandra V. Gavshina
collection DOAJ
description Abstract Wild-type SAASoti and its monomeric variant mSAASoti can undergo phototransformations, including reversible photoswitching of the green form to a nonfluorescent state and irreversible green-to-red photoconversion. In this study, we extend the photochemistry of mSAASoti variants to enable reversible photoswitching of the red form. This result is achieved by rational and site-saturated mutagenesis of the M163 and F177 residues. In the case of mSAASoti it is M163T substitution that leads to the fastest switching and the most photostable variant, and reversible photoswitching can be observed for both green and red forms when expressed in eukaryotic cells. We obtained a 13-fold increase in the switching efficiency with the maximum switching contrast of the green form and the appearance of comparable switching of the red form for the C21N/M163T mSAASoti variant. The crystal structure of the C21N mSAASoti in its green on-state was obtained for the first time at 3.0 Å resolution, and it is in good agreement with previously calculated 3D-model. Dynamic network analysis reveals that efficient photoswitching occurs if motions of the 66H residue and phenyl fragment of chromophore are correlated and these moieties belong to the same community.
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spelling doaj.art-2e42d3f72ccc4b7094b8c0cb56b72db62024-04-21T11:19:21ZengNature PortfolioScientific Reports2045-23222024-04-0114111210.1038/s41598-024-59364-1The role of the correlated motion(s) of the chromophore in photoswitching of green and red forms of the photoconvertible fluorescent protein mSAASotiAlexandra V. Gavshina0Ilya D. Solovyev1Maria G. Khrenova2Konstantin M. Boyko3Larisa A. Varfolomeeva4Mikhail E. Minyaev5Vladimir O. Popov6Alexander P. Savitsky7A.N. Bach Institute of Biochemistry, Federal Research Centre ‘Fundamentals of Biotechnology’ of the Russian Academy of SciencesA.N. Bach Institute of Biochemistry, Federal Research Centre ‘Fundamentals of Biotechnology’ of the Russian Academy of SciencesA.N. Bach Institute of Biochemistry, Federal Research Centre ‘Fundamentals of Biotechnology’ of the Russian Academy of SciencesA.N. Bach Institute of Biochemistry, Federal Research Centre ‘Fundamentals of Biotechnology’ of the Russian Academy of SciencesA.N. Bach Institute of Biochemistry, Federal Research Centre ‘Fundamentals of Biotechnology’ of the Russian Academy of SciencesN.D. Zelinsky Institute of Organic Chemistry Russian Academy of SciencesA.N. Bach Institute of Biochemistry, Federal Research Centre ‘Fundamentals of Biotechnology’ of the Russian Academy of SciencesA.N. Bach Institute of Biochemistry, Federal Research Centre ‘Fundamentals of Biotechnology’ of the Russian Academy of SciencesAbstract Wild-type SAASoti and its monomeric variant mSAASoti can undergo phototransformations, including reversible photoswitching of the green form to a nonfluorescent state and irreversible green-to-red photoconversion. In this study, we extend the photochemistry of mSAASoti variants to enable reversible photoswitching of the red form. This result is achieved by rational and site-saturated mutagenesis of the M163 and F177 residues. In the case of mSAASoti it is M163T substitution that leads to the fastest switching and the most photostable variant, and reversible photoswitching can be observed for both green and red forms when expressed in eukaryotic cells. We obtained a 13-fold increase in the switching efficiency with the maximum switching contrast of the green form and the appearance of comparable switching of the red form for the C21N/M163T mSAASoti variant. The crystal structure of the C21N mSAASoti in its green on-state was obtained for the first time at 3.0 Å resolution, and it is in good agreement with previously calculated 3D-model. Dynamic network analysis reveals that efficient photoswitching occurs if motions of the 66H residue and phenyl fragment of chromophore are correlated and these moieties belong to the same community.https://doi.org/10.1038/s41598-024-59364-1Biphotochromic fluorescent proteinsGreen-to-red photoconversionReversible photoswitchingPhotochemistry of fluorescent proteinsCrystal structureDynamic network analysis
spellingShingle Alexandra V. Gavshina
Ilya D. Solovyev
Maria G. Khrenova
Konstantin M. Boyko
Larisa A. Varfolomeeva
Mikhail E. Minyaev
Vladimir O. Popov
Alexander P. Savitsky
The role of the correlated motion(s) of the chromophore in photoswitching of green and red forms of the photoconvertible fluorescent protein mSAASoti
Scientific Reports
Biphotochromic fluorescent proteins
Green-to-red photoconversion
Reversible photoswitching
Photochemistry of fluorescent proteins
Crystal structure
Dynamic network analysis
title The role of the correlated motion(s) of the chromophore in photoswitching of green and red forms of the photoconvertible fluorescent protein mSAASoti
title_full The role of the correlated motion(s) of the chromophore in photoswitching of green and red forms of the photoconvertible fluorescent protein mSAASoti
title_fullStr The role of the correlated motion(s) of the chromophore in photoswitching of green and red forms of the photoconvertible fluorescent protein mSAASoti
title_full_unstemmed The role of the correlated motion(s) of the chromophore in photoswitching of green and red forms of the photoconvertible fluorescent protein mSAASoti
title_short The role of the correlated motion(s) of the chromophore in photoswitching of green and red forms of the photoconvertible fluorescent protein mSAASoti
title_sort role of the correlated motion s of the chromophore in photoswitching of green and red forms of the photoconvertible fluorescent protein msaasoti
topic Biphotochromic fluorescent proteins
Green-to-red photoconversion
Reversible photoswitching
Photochemistry of fluorescent proteins
Crystal structure
Dynamic network analysis
url https://doi.org/10.1038/s41598-024-59364-1
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