Difference between revisions of "Part:BBa K1936000"

Revision as of 18:45, 15 October 2016

ePDZ-mCherry-BAX184E

ALERT !!!Under construction !!

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 utilized the binding of ePDZ to the J-alpha-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 inner membrane space to the cytosol which leads to apoptosome formation and therefore triggers cell death. The BaxS184E mutant has been shown to not migrate from cytosol to OMM on its own. To induce apoptosis by expressing BaxS184E recruitment to the OMM is necessary. [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