Part:BBa_K1936000

ePDZ-mCherry-BAX184E

Usage and Biology

PDZ domains are commonly found in signaling proteins and play important roles in recruitment and scaffold association. The engineered PDZ (ePDZ) domain is designed to interact with LOV2 peptide (blue light inducible system). We used the binding capability of ePDZ to the Jα helix of LOV2 to recruit ePDZ-mCherry-BaxS184E fusion protein to the outer mitochondrial membrane (OMM) (BBa_K1936001). [1]

The human Bax protein acts as cofactor for the tumor suppressor gene p53 that is associated with acceleration of apoptosis. It translocates from the cytosol to the outer mitochondrial membrane (OMM) where it participates in the formation of mitochondrial apoptosis-induced channels (MAC). Through those channels cytochrome c can be released from the mitochondrial intermembrane space to the cytosol which leads to apoptosome formation and therefore triggers cell death. The BaxS184E mutant has been shown not to migrate from cytosol to OMM autonomously. Induction of apoptosis by BaxS184E requires its recruitment to the OMM. [2]

The red fluorescent protein mCherry is used to track the fusion protein localization inside a cell by microscopy.

References

[1] Konrad Müller, Raphael Engesser, Jens Timmer, Matias D. Zurbriggen, and Wilfried Weber (2014) Orthogonal Optogenetic Triple-Gene Control in Mammalian Cells. ACS Synth Biol. 2014 Nov 21;3(11):796-801. doi: 10.1021/sb500305v. Epub 2014 Oct 28.

[2] Hughes RM, Freeman DJ, Lamb KN, Pollet RM, Smith WJ, Lawrence DS. Optogenetic apoptosis: light-triggered cell death. Angew Chem Int Ed Engl. 2015 Oct 5;54(41):12064-8. doi: 10.1002/anie.201506346. Epub 2015 Aug 25.

Sequence and Features

Assembly Compatibility:

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

  Illegal BglII site found at 1327
  Illegal BamHI site found at 1796
  Illegal XhoI site found at 1331
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]