Part:BBa_K5403021

This composite parts consists of GFP with a Spytag at its N-terminus and a hexahistidine tag (His-Tag) at its C-terminus. GFP and Spytag are joined via a flexible linker (BBa_K5403002), to avoid that the conjugation of these two would cause misfolding.

Spytag is a peptide that specifically binds Spycatcher. In Figure 1, Spycatcher is shown in blue in Spytag is shown in orange. Conjugating Spytag to a protein of interest can have several applications. For example, it allows one to gain control over the localization of the protein of interest. It can also function to determine the location of Spycatcher. Additionally, it may be used to colocalize the protein of interest with another protein which is recombinantly expressed to fuse with Spycatcher.

Figure 1: complex of SpyCatcher (green) and SpyTag (orange) (Li et al., 2014).

References

Li, L., Fierer, J. O., Rapoport, T. A., & Howarth, M. (2014). Structural Analysis and Optimization of the Covalent Association between SpyCatcher and a Peptide Tag. Journal Of Molecular Biology, 426(2), 309–317. https://doi.org/10.1016/j.jmb.2013.10.021