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Novel Phototransformable Fluorescent Protein SAASoti with Unique Photochemical Properties

1
Department of Chemistry, M.V. Lomonosov Moscow State University, Leninskie Gory 1/3, Moscow 119991, Russia
2
A.N. Bach Institute of Biochemistry, Research Center of Biotechnology of the Russian Academy of Sciences, Leninsky Ave. 33, bld. 2, Moscow 119071, Russia
*
Author to whom correspondence should be addressed.
Int. J. Mol. Sci. 2019, 20(14), 3399; https://doi.org/10.3390/ijms20143399
Received: 14 June 2019 / Revised: 4 July 2019 / Accepted: 8 July 2019 / Published: 11 July 2019
(This article belongs to the Special Issue Imaging with Fluorescent Proteins)
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Abstract

SAASoti is a unique fluorescent protein (FP) that combines properties of green-to-red photoconversion and reversible photoswitching (in its green state), without any amino acid substitutions in the wild type gene. In the present work, we investigated its ability to photoswitch between fluorescent red (‘on’) and dark (‘off’) states. Surprisingly, generated by 400 nm exposure, the red form of SAASoti (R1) does not exhibit any reversible photoswitching behavior under 550 nm illumination, while a combination of prior 470 nm and subsequent 400 nm irradiation led to the appearance of another—R2—form that can be partially photoswitched (550 nm) to the dark state, with a very fast recovery time. The phenomenon might be explained by chemical modification in the chromophore microenvironment during prior 470 nm exposure, and the resulting R2 SAASoti differs chemically from the R1 form. The suggestion is supported by the mass spectrometry analysis of the tryptic peptides before and after 470 nm light exposure, that revealed Met164 oxidation, as proceeds in another dual phototransformable FP, IrisFP. View Full-Text
Keywords: fluorescent proteins; photoconversions of fluorescent proteins; reversible photoswitching fluorescent proteins; photoconversions of fluorescent proteins; reversible photoswitching
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).

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Solovyev, I.D.; Gavshina, A.V.; Savitsky, A.P. Novel Phototransformable Fluorescent Protein SAASoti with Unique Photochemical Properties. Int. J. Mol. Sci. 2019, 20, 3399.

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