Part:BBa_K3064016

mmuIgG-Fc->HA-Trim21

With this composite part, IgG-Fc and Trim21 can be expressed in cells, conducing to an improvement of Trim-Away method for the degradation of target protein. Also, HA Tag was added to N-terminal of Trim21, making it easier for Trim21 to be detected by Western Blotting.

Usage and Biology

Immunoglobulin G (IgG) is a type of antibody, of which fragment crystallizable region (Fc) is the tail region. Trim21, an E3 ubiquitin-protein ligase functioning in the intracellular antibody-mediated proteolysis pathway, binds with high affinity to the Fc domain of antibody¹ .

If this part is linked with the target protein recognition module (such as the nanobody in our 2018 project and glucagon in our 2019 project), this recombinant can be used for protein degradation. Here is the brief mechanism:

• The target protein recognition module recognizes and binds to the target protein

• Fc recruits Trim21

• The target protein is ubiquitinated and finally degraded by 26S proteasome

Sequence and Features

Assembly Compatibility:

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

  Illegal BamHI site found at 646
  Illegal BamHI site found at 1349
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 747
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 7
  

References

1.James LC, Keeble AH, Khan Z, Rhodes DA, Trowsdale J. "Structural basis for PRYSPRY-mediated tripartite motif (TRIM) protein function". Proceedings of the National Academy of Sciences of the United States of America. 104 (15): 6200–5. Bibcode:2007PNAS..104.6200J.(2007)