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Design of AsLOV2 domain as a carrier of light-induced dissociable FMN photosensitizer. | LitMetric

AI Article Synopsis

  • Flavin mononucleotide (FMN) is a strong photosensitizer that produces singlet oxygen effectively, but its efficiency drops when it's incorporated into protein structures.
  • The study explores a method to enhance FMN's oxygen production by engineering protein interactions, specifically inducing FMN release through light-triggered oxidation of cysteine residues.
  • Using different variants of the LOV2 domain from Avena sativa, the research shows that improving FMN release correlates with increased singlet oxygen production, allowing for better application of FMN in genetically encoded photosensitizers.

Article Abstract

Flavin mononucleotide (FMN) is a highly efficient photosensitizer (PS) yielding singlet oxygen ( O ). However, its O production efficiency significantly decreases upon isoalloxazine ring encapsulation into the protein matrix in genetically encoded photosensitizers (GEPS). Reducing isoalloxazine ring interactions with surrounding amino acids by protein engineering may increase O production efficiency GEPS, but at the same time weakened native FMN-protein interactions may cause undesirable FMN dissociation. Here, in contrast, we intentionally induce the FMN release by light-triggered sulfur oxidation of strategically placed cysteines (oxidation-prone amino acids) in the isoalloxazine-binding site due to significantly increased volume of the cysteinyl side residue(s). As a proof of concept, in three variants of the LOV2 domain of Avena sativa (AsLOV2), namely V416C, T418C, and V416C/T418C, the effective O production strongly correlated with the efficiency of irradiation-induced FMN dissociation (wild type (WT) < V416C < T418C < V416C/T418C). This alternative approach enables us: (i) to overcome the low O production efficiency of flavin-based GEPSs without affecting native isoalloxazine ring-protein interactions and (ii) to utilize AsLOV2, due to its inherent binding propensity to FMN, as a PS vehicle, which is released at a target by light irradiation.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10949324PMC
http://dx.doi.org/10.1002/pro.4921DOI Listing

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