Flavin-tag: A Facile Method for Site-Specific Labeling of Proteins with a Flavin Fluorophore.
Yapei TongMisun LeeJeroen DrenthMarco W FraaijePublished in: Bioconjugate chemistry (2021)
Site-specific protein labeling methods are highly valuable tools for research and applications. We present a new protein labeling method that allows covalent attachment of a chromo- and fluorogenic flavin (FMN) to any targeted protein using a short flavinylation peptide-tag. We show that this peptide can be as short as 7 residues and can be located at the N-terminus, C-terminus, or in internal regions of the target protein. Analogous to kinase-catalyzed phosphorylation, the flavin is covalently attached via a stable phosphothreonyl linkage. The site-specific covalent tethering of FMN is accomplished by using a bacterial flavin transferase. The covalent coupling of FMN was shown to work in Escherichia coli and Saccharomyces cerevisiae cells and could be performed in vitro, rendering the "Flavin-tag" method a powerful tool for the selective decoration of proteins with a biocompatible redox-active fluorescent chromophore.
Keyphrases
- escherichia coli
- saccharomyces cerevisiae
- protein protein
- amino acid
- induced apoptosis
- quantum dots
- binding protein
- room temperature
- oxidative stress
- staphylococcus aureus
- drug delivery
- cancer therapy
- endoplasmic reticulum stress
- cell cycle arrest
- protein kinase
- hepatitis c virus
- hiv infected
- cystic fibrosis
- biofilm formation
- hiv testing
- fluorescent probe