Novel Phototransformable Fluorescent Protein SAASoti with Unique Photochemical Properties

International Journal of Molecular Sciences
Ilya D SolovyevAlexander P Savitsky

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.

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Citations

Jun 5, 2021·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Elena A ProtasovaAlexey M Bogdanov
May 29, 2020·Journal of the American Chemical Society·Elke De ZitterDominique Bourgeois

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Methods Mentioned

BETA
irradiating
Fluorescence

Software Mentioned

Maple
FlexAnalysis
SAASoti
pcDronpa
Biotools
Origin
NijiFP
Mascot

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