Part:BBa_K4040020

Synthetic Receptor GP130-TEV

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 1508
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

Based on the mechanism of Tango/MESA, we bind TEV-protase to GP130. The MESA (modular extracellular sensor architecture) system is based on bringing intracellular membrane-anchored TEV (tobacco etch virus) protease into proximity to a membrane-anchored transcription factor in response to extracellular ligand binding. This is followed by cleavage and nuclear localization of the transcription factor and has been used to sense human VEGF (vascular endothelial growth factor)[1]. We used CMV as the promoter. When the concentration of IL-6 rises to a certain level, IL-6 binds to mIL-6R (membrane-bound IL-6R), homodimerization of gp130 is induced. A high-affinity functional receptor complex of IL-6, IL-6R and gp130 is formed.  Then, TEV-protase connected behind is activated. TEV-protase binds to TCS and releases transcription factors on the other two receptors through enzyme digestion to initiate the subsequent pathway.  

Figure. 1The structure of gp130-TEV.

Figure. 2The structure and downstream activation of the composite IL-6R.

References

[1]Schwarz, K. A., Daringer, N. M., Dolberg, T. B. & Leonard, J. N. Rewiring human cellular input-output using modular extracellular sensors. Nat. Chem. Biol. 13, 202–209 (2017).