Peptide interactions are nearly always weak and reversible, limiting application of peptides for assembly and control of biomolecules. We have designed tools enabling irreversible peptide interaction, based on a special feature of the chemistry of the human pathogen Streptococcus pyogenes. By rational engineering of a protein from this bacterium involved with host invasion, we obtained a peptide tag (SpyTag) bearing a carboxylic acid side-chain which spontaneously reacts with an amine side-chain, to form an amide bond locking the peptide to the protein partner (SpyCatcher). This reaction is rapid, high-yielding, genetically encodable and specific on cells. Complementation of this split protein has been analyzed by NMR and advanced by rational and library-based engineering. Applications of this protein padlock will be presented to show new possibilities for mammalian cell imaging, for enhancing enzyme function, and for controlling force within living cells, such as at chromosome segregation.

From flesh-eating bacteria to protein superglue

245th National Meeting (2013)

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