TY  - JOUR
TI  - Detection of Membrane Protein Interactions by Cell-based Tango Assays
AU  - Yan, Yan 
AU  - Xu, Ting-Hai 
AU  - Harikumar, Kaleeckal G.
AU  - Miller, Laurence J.
AU  - Melcher, Karsten 
AU  - Xu, H. Eric
VL  - 7
IS  - 22
PY  - 2017
DA  - 2017/11/20
SP  - e2903
C1  - Bio-protocol 2017;7:e2903
DO  - 10.21769/BioProtoc.2903
UR  - https://doi.org/10.21769/BioProtoc.2903
AB  - The Tango assay is a protein-protein interaction assay, in which a transcription factor (rTA) is fused to a membrane-bound protein via a linker that contains a cleavage site for TEV protease, whereas a soluble interaction partner is fused to TEV protease (Barnea et al., 2008). Association between the two interaction partners leads to an efficient cleavage of the transcription factor, allowing it to translocate to the nucleus and activate a luciferase reporter gene as measurement of the interactions. In this modified assay, we fused one copy of the membrane-spanning amyloid precursor protein (APP) C99 region to TEV site-rTA (C99-TEV site-rTA) and a second copy to TEV protease (C99-TEV) to analyze intramembrane C99-C99 interaction in live cells.
KW  - Tango assay
KW  - γ-Secretase
KW  - Cleavage
KW  - Activity
KW  - Interaction
KW  - C99
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False