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. 2016 Oct-Dec;8(4):118-123.

Flavoprotein miniSOG Cytotoxisity Can Be Induced By Bioluminescence Resonance Energy Transfer

E I Shramova  1 , G M Proshkina  1 , S P Chumakov  1 , Yu M Khodarovich  1 , S M Deyev  2
Affiliations

Flavoprotein miniSOG Cytotoxisity Can Be Induced By Bioluminescence Resonance Energy Transfer

E I Shramova et al. Acta Naturae. 2016 Oct-Dec.

Abstract

In this study, we investigated the possibility of phototoxic flavoprotein miniSOG (photosensitizer) excitation in cancer cells by bioluminescence occurring when luciferase NanoLuc oxidizes its substrate, furimazine. We have shown that the phototoxic flavoprotein miniSOG expressed in eukaryotic cells in fusion with NanoLuc luciferase is activated in the presence of its substrate, furimazine. Upon such condition, miniSOG possesses photoinduced cytotoxicity and causes a 48% cell death level in a stably transfected cell line.

Keywords: NanoLuc luciferase; bioluminescence resonance energy transfer; flavoprotein miniSOG; photodynamic therapy.

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Figures

Fig. 1
Fig. 1
Bioluminescence system based on luciferase, furimazine, and miniSOG. A – Schematic illustration of the BRET system for PDT. B – Gene construct encoding NanoLuc, peptide linker GGGGS and cytotoxic module miniSOG within one reading frame. C – Normalized emission spectrum of furimamide (NanoLucem) and normalized excitation (miniSOGex) and emission (miniSOGem) spectra of miniSOG.
Fig. 2
Fig. 2
Emission spectra of the bioluminescence systems NanoLuc-furimazine (NanoLuc) and NanoLuc-furimazine-miniSOG (NanoLuc-miniSOG) in the presence of FMN.
Fig. 3
Fig. 3
Functional characteristics of the bioluminescence system NanoLuc-furimazine-miniSOG. A – Dependence of NanoLuc-miniSOG emission spectra on furimazine and FMN concentrations. B – Dependence of the cytotoxic effect of the NanoLuc-miniSOG fusion protein on the furimazine concentration.
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