Isopeptag

Isopeptag is a 16 amino acid peptide tag (TDKDMTITFTNKKDAE) that can be genetically linked to proteins without interfering with protein folding.[1] What makes the isopeptag different from other peptide tags is that it can bind its binding protein through a permanent and irreversible covalent bond. Other peptide tags generally bind their targets through weak non-covalent interactions, thus limiting their use in applications where molecules experience extreme forces. The isopeptags covalent binding to its target overcomes these barriers and allows target proteins to be studied in harsher molecular environments.

Development

The isopeptag was developed by dissecting the pilin protein (Spy0128) from Streptococcus pyogenes. Spy0128 contains two intramolecular isopeptide bonds,[2] and to generate the isopeptag one of these bonds was split by removing the last β-sheet in the protein.

Mode of Action

When the isopeptag is bound to a target protein, it spontaneously binds its binding partner through an isopeptide bond. The reaction is robust and occurs at various temperatures from 4-37°C, a pH range of 5-8, and in the presence of commonly used detergents. Also, the reaction is independent of the redox state of the environment and can occur equally well in both reducing and oxidizing conditions.[1]

Applications

The covalent binding of the isopeptag to its binding partner can be used to permanently link proteins together in the complex environment of a bacterial cell, to target proteins of interest for cellular imaging, and to develop new protein structures.[1]

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gollark: Which is probably unnecessary and results in security issues and bigger datas.
gollark: You can include some kind of JSON schema (because there are lots of things for that) with the docs. Though presumably someone will notice if the API is missing a field they need.
gollark: Your API should have documentation. Just use that.
gollark: The best format for data exchange between APIs is of course small SQLite databases.

References
  1. Zakeri, Bijan; Howarth, Mark (7 April 2010). "Spontaneous Intermolecular Amide Bond Formation between Side Chains for Irreversible Peptide Targeting". Journal of the American Chemical Society. 132 (13): 4526–4527. doi:10.1021/ja910795a. PMID 20235501.
  2. Kang,H.J., Coulibaly,F., Clow,F., Proft,T., and Baker,E.N. (2007). Stabilizing isopeptide bonds revealed in gram-positive bacterial pilus structure. Science 318, 1625-1628.
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