Part:BBa_K4247009:Design
SnoopCatcher
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
In our project, the goal of SnoopTag/Catcher was to specifically bind mussel foot protein 151 (composite part BBa_K247021) to a set of minispidroins (composite parts BBa_K4247010 and BBa_K4247013). To build our system we:
1) Synthesised SnoopCatcher as part of the second half of mussel foot protein 151 (part BBa_K247017) at the N-terminus and using the linker GGGSG. The SnoopCatcher protein sequence is KPLRGAVFSLQKQHPDYPDIYGAIDQNGTYQNVRTGEDGKLTFKNLSDGKYRLFENSEPAGYKPVQNKPIVAFQIVNGEVRDVTSIVPQDIPATYEFTNGKHYITNEPIPPK
2) SnoopTag was linked to our protein of interest at the C-terminus via a user PCR reaction, because the N-terminus already had the 6x His-tag. Nonetheless, it seems to be generally more common to link SnoopTag to the N-terminus of the desired protein (based on the SpyBank database of Howarth group). The SnoopTag protein sequence is KLGDIEFIKVNKPPK
Source
This sequence was taken from the SpyBank database which can be accessed through the link: http://www.howarthgroup.org/info
References
Veggiani G, Nakamura T, Brenner MD, Gayet RV, Yan J, Robinson CV, Howarth M.Programmable polyproteams built using twin peptide superglues. Proc Natl Acad Sci U S A. 2016 Feb 2;113(5):1202-7.
Tan LL, Hoon SS, Wong FT. Kinetic Controlled Tag-Catcher Interactions for Directed Covalent Protein Assembly. PLoS ONE 11(10): e0165074.
Hatlem D, Trunk T, Linke D, Leo JC. Catching a SPY: Using the SpyCatcher-SpyTag and Related Systems for Labeling and Localizing Bacterial Proteins. Int J Mol Sci. 2019 Apr 30;20(9):2129. doi: 10.3390/ijms20092129. PMID: 31052154; PMCID: PMC6539128.