Light induces oxidative damage and protein stability in the fungal photoreceptor Vivid

PloS One
Carmen Noemí Hernández-CandiaBraulio Gutiérrez-Medina

Abstract

Flavin-binding photoreceptor proteins sense blue-light (BL) in diverse organisms and have become core elements in recent optogenetic applications. The light-oxygen-voltage (LOV) protein Vivid (VVD) from the filamentous fungus Neurospora crassa is a classic BL photoreceptor, characterized by effecting a photocycle based on light-driven formation and subsequent spontaneous decay of a flavin-cysteinyl adduct. Here we report that VVD presents alternative outcomes to light exposure that result in protein self-oxidation and, unexpectedly, rise of stability through kinetic control. Using optical absorbance and mass spectrometry we show that purified VVD develops amorphous aggregates with the presence of oxidized residues located at the cofactor binding pocket. Light exposure increases oxidative levels in VVD and specific probe analysis identifies singlet oxygen production by the flavin. These results indicate that VVD acts alternatively as a photosensitizer, inducing self-oxidative damage and subsequent aggregation. Surprisingly, BL illumination has an additional, opposite effect in VVD. We show that light-induced adduct formation establishes a stable state, delaying protein aggregation until photoadduct decay occurs. In accordance, r...Continue Reading

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Citations

Oct 28, 2019·Journal of Mass Spectrometry : JMS·Emerson Finco MarquesPaolo Di Mascio
Feb 10, 2021·Biochimica Et Biophysica Acta. Proteins and Proteomics·Braulio Gutiérrez-Medina, Carmen Noemí Hernández-Candia
Nov 18, 2020·BMC Biology·Mikael MolinAnders Blomberg
Dec 24, 2021·Nature Chemical Biology·William BenmanLukasz J Bugaj

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Methods Mentioned

BETA
electrophoresis
Transmission electron microscopy
Fluorescence
fluorescence imaging

Software Mentioned

Mathematica
Protein Lynx Global Server
MASCOT

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