Part:BBa_K3765001
SpyCatcher002
115 amino acid protein partner to SpyTag/SpyTag002 [1], with which it forms an isopeptide bond, used for anchoring two fusion proteins.
Usage and Biology
Streptococcus pyogenes has a fibronectin binding protein, FbaB. The collagen adhesin domain (CnaB2) of FbaB contains an internal isopeptide bond between Lys31 and Asp117 that confers exceptional stability. Zakeri et al. (PNAS, 2012) split the CnaB2 domain into an N-terminal protein fragment (SpyCatcher), containing Lys31, and a C-terminal peptide of 13 amino acids (SpyTag), containing Asp117. These parts spontaneously reconstitute to form an isopeptide bond under a wide range of pH values, temperatures, and redox conditions and in various detergents, remaining robust despite boiling or forces exceeding 1 nN, a breakage force at which streptavidin–biotin attachments and antibody–antigen interactions fail. In 2017, Keeble et al. (Angew. Chem. Int. Ed., 2017) reported SpyCatcher002 by using phage display and selecting for accelerated amidation and increased cross-reactivity with SpyTag, while simultaneously ensuring lower self-reactivity to prevent the formation of SpyCatcher-SpyCatcher dimer. Thermostability of SpyCatcher002 is comparable with SpyCatcher.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal XhoI site found at 150
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 235
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