Tyrosinase(TYR) is an important polyphenolic oxidase enzyme and usually regards as a biomarker of melano ma cancer.Highly effective tracking TYR activity in vivo will help to study the mechanism of TYR in living organ...Tyrosinase(TYR) is an important polyphenolic oxidase enzyme and usually regards as a biomarker of melano ma cancer.Highly effective tracking TYR activity in vivo will help to study the mechanism of TYR in living organisms and forecasts related diseases.In this study,we present a novel TYR-activatable fluo rescent probe(CHMC-DOPA) for tracking TYR activity in vitro and in vivo.CHMC-DOPA is constructed by incorporating dopamine(DOPA) moiety into a fluorescent chloro-hydroxyl-merocyanine(CHMC)scaffold.Upon exposure to TYR,the dopamine unit in CHMC-DOPA is oxidized to a dopaquinone derivative,and an intramolecular photo-induced electron transfer(PET) process between CHMC fluorophore and o-dopaquinone will take place,the fluorescence of CHMC-DOPA is quenched rapidly.Therefore,the evaluation of TYR activity is established in terms of the relationship between fluorescence quenching efficiency and TYR activity.In our experiments,CHMC-DOPA shows various advantages,such as fast response(8 min),low concentration of TYR activation(0.5 U/mL),good water-solubility,as well as the lowest detection limit(0.003 U/mL) compared with previously reported works.Furthermore,CHMCDO PA also exhibits excellent cell membrane permeability and low cytotoxicity,which is successfully used to monitor endogenous TYR activity in living cancer cells and zebrafish models.CHMC-DOPA performs well,and we anticipate that this newly designed novel platform will provide an alternative for high effective monitoring TYR activity in biosystems.展开更多
基金We gratefully acknowledge the financial support by the National Natural Science Foundation of China(Nos.21702053,21901066,21676075,51603064)Natural Science Foundation of Hubei Province(No.2020CFB601)+1 种基金Outstanding Young and Middle-aged Scientific Innovation Team of Colleges and Universities of Hubei Province(No.T201908)partially supported by Open Funding Project of the State Key Laboratory of Biocatalysis and Enzyme Engineering.
文摘Tyrosinase(TYR) is an important polyphenolic oxidase enzyme and usually regards as a biomarker of melano ma cancer.Highly effective tracking TYR activity in vivo will help to study the mechanism of TYR in living organisms and forecasts related diseases.In this study,we present a novel TYR-activatable fluo rescent probe(CHMC-DOPA) for tracking TYR activity in vitro and in vivo.CHMC-DOPA is constructed by incorporating dopamine(DOPA) moiety into a fluorescent chloro-hydroxyl-merocyanine(CHMC)scaffold.Upon exposure to TYR,the dopamine unit in CHMC-DOPA is oxidized to a dopaquinone derivative,and an intramolecular photo-induced electron transfer(PET) process between CHMC fluorophore and o-dopaquinone will take place,the fluorescence of CHMC-DOPA is quenched rapidly.Therefore,the evaluation of TYR activity is established in terms of the relationship between fluorescence quenching efficiency and TYR activity.In our experiments,CHMC-DOPA shows various advantages,such as fast response(8 min),low concentration of TYR activation(0.5 U/mL),good water-solubility,as well as the lowest detection limit(0.003 U/mL) compared with previously reported works.Furthermore,CHMCDO PA also exhibits excellent cell membrane permeability and low cytotoxicity,which is successfully used to monitor endogenous TYR activity in living cancer cells and zebrafish models.CHMC-DOPA performs well,and we anticipate that this newly designed novel platform will provide an alternative for high effective monitoring TYR activity in biosystems.
