Part:BBa_K2721027:Design

LDH with SdyTag and SpyCatcher

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 43
  Illegal AgeI site found at 79
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 976
  

Design Notes

The tag-catcher pairs are ideal for binding, immobilization and creating new kinds of protein architectures. Our team use them as scaffold of protein assembly. LDH is one of the enzymes fixed on the scaffold to catalyze relevant chemical reaction.

We have mutated some illegal enzyme sites through synonymous substitutions to make it compatible with RFC[10].

Source

SpyTag/SpyCatcher is derived from Streptococcus pyogenes fibronectin-binding protein. SdyTag/SdyCatcher is derived from a related fibronectin-binding protein in Streptococcus dysgalactiae. The gene sequence of LDH was isolated from E. coli.

References

1. Tan LL, Hoon SS, Wong FT. Kinetic Controlled Tag-Catcher Interactions for Directed Covalent Protein Assembly. PLoS ONE 11(10): e0165074. https://doi.org/10.1371/journal.pone.0165074

2. Zakeri B, Fierer JO, Celik E, Chittock EC, Schwarz-Linek U, Moy VT, Howarth M. Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin. Proc Natl Acad Sci U S A. 2012 Mar 20;109(12):E690-7.