1. Flavin-Tag
- Author
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Jeroen Drenth, Misun Lee, Yapei Tong, Marco W. Fraaije, and Biotechnology
- Subjects
Saccharomyces cerevisiae Proteins ,Fluorophore ,Stereochemistry ,Saccharomyces cerevisiae ,Biomedical Engineering ,Pharmaceutical Science ,Bioengineering ,Peptide ,Flavin group ,chemistry.chemical_compound ,Flavins ,Escherichia coli ,Transferase ,heterocyclic compounds ,Phosphorylation ,Fluorescent Dyes ,Pharmacology ,chemistry.chemical_classification ,Staining and Labeling ,biology ,Escherichia coli Proteins ,Organic Chemistry ,Proteins ,biology.organism_classification ,Fluorescence ,chemistry ,Covalent bond ,Target protein ,Biotechnology - Abstract
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.
- Published
- 2021