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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Nature Portfolio
2024-04-01
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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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