Genetically encoding thyronine for fluorescent detection of peroxynitrite.
Bioorg Med Chem
; 28(18): 115665, 2020 09 15.
Article
in En
| MEDLINE
| ID: mdl-32828428
ABSTRACT
Peroxynitrite is a highly reactive oxidant effecting cell signaling and cell death. Here we report a fluorescent protein probe to selectively detect peroxynitrite. A novel unnatural amino acid, thyronine (Thy), was genetically encoded in E. coli and mammalian cells by evolving an orthogonal tRNAPyl/ThyRS pair. Incorporation of Thy into the chromophore of sfGFP or cpsGFP afforded a virtually non-fluorescent reporter. Upon treatment with peroxynitrite, Thy was converted into tyrosine via O-dearylation, regenerating GFP fluorescence in a time- and concentration-dependent manner. Genetically encoded thyronine may also be valuable for other redox applications.
Key words
Full text:
1
Database:
MEDLINE
Main subject:
Thyronines
/
Peroxynitrous Acid
/
Green Fluorescent Proteins
/
Fluorescent Dyes
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
En
Year:
2020
Type:
Article