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

0584828 - MBÚ 2025 RIV US eng J - Článek v odborném periodiku
Felčíková, K. - Hovan, A. - Polák, Marek - Loginov, Dmitry Sergej - Holotová, V. - Diaz, C. - Kožář, T. - Lee, O. - Varhač, R. - Novák, Petr - Bánó, G. - Sedlák, E.
Design of AsLOV2 domain as a carrier of light-induced dissociable FMN photosensitizer.
Protein Science. Roč. 33, č. 4 (2024), č. článku e4921. ISSN 0961-8368. E-ISSN 1469-896X
GRANT EU: European Commission(XE) 731077 - EU_FT-ICR_MS
Výzkumná infrastruktura: CIISB II - 90127
Institucionální podpora: RVO:61388971
Klíčová slova: singlet oxygen generation * molecular-dynamics * rational design * simulation * efficient * ewald * flavin cofactor * genetically encoded photosensitizers * LOV2 domain * miniSOG * singlet oxygen
Obor OECD: Microbiology
Impakt faktor: 8, rok: 2022
Způsob publikování: Omezený přístup
https://onlinelibrary.wiley.com/doi/epdf/10.1002/pro.4921

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.
Trvalý link: https://hdl.handle.net/11104/0352674