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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F24%3A00584828" target="_blank" >RIV/61388971:_____/24:00584828 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/00216224:90127/24:00139163 RIV/00216208:11310/24:10478173

Výsledek na webu

<a href="https://onlinelibrary.wiley.com/doi/epdf/10.1002/pro.4921" target="_blank" >https://onlinelibrary.wiley.com/doi/epdf/10.1002/pro.4921</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1002/pro.4921" target="_blank" >10.1002/pro.4921</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10608 - Biochemistry and molecular biology

Projekt

<a href="/cs/project/LX22NPO5107" target="_blank" >LX22NPO5107: Národní ústav pro neurologický výzkum</a><br>

Návaznosti

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Protein Science

ISSN

0961-8368

e-ISSN

1469-896X

Svazek periodika

33

Číslo periodika v rámci svazku

4

Stát vydavatele periodika

US - Spojené státy americké

Počet stran výsledku

15

Strana od-do

e4921

Kód UT WoS článku

001187153900001

EID výsledku v databázi Scopus

2-s2.0-85188256683