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Design of AsLOV2 domain as a carrier of light-induced dissociable FMN photosensitizer
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SYSNO ASEP 0584828 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Design of AsLOV2 domain as a carrier of light-induced dissociable FMN photosensitizer Author(s) Felčíková, K. (SK)
Hovan, A. (SK)
Polák, Marek (MBU-M)
Loginov, Dmitry Sergej (MBU-M) RID
Holotová, V. (SK)
Diaz, C. (SK)
Kožář, T. (SK)
Lee, O. (SK)
Varhač, R. (SK)
Novák, Petr (MBU-M) RID, ORCID
Bánó, G. (SK)
Sedlák, E. (SK)Article number e4921 Source Title Protein Science. - : Wiley - ISSN 0961-8368
Roč. 33, č. 4 (2024)Language eng - English Country US - United States Keywords singlet oxygen generation ; molecular-dynamics ; rational design ; simulation ; efficient ; ewald ; flavin cofactor ; genetically encoded photosensitizers ; LOV2 domain ; miniSOG ; singlet oxygen OECD category Microbiology Research Infrastructure CIISB II - 90127 - Masarykova univerzita Method of publishing Limited access Institutional support MBU-M - RVO:61388971 UT WOS 001187153900001 EID SCOPUS 85188256683 DOI 10.1002/pro.4921 Annotation Flavin mononucleotide (FMN) is a highly efficient photosensitizer (PS) yielding singlet oxygen (O-1(2)). However, its O-1(2) 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 1O2 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-1(2) 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-1(2) 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. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2025 Electronic address https://onlinelibrary.wiley.com/doi/epdf/10.1002/pro.4921
Number of the records: 1