AKT1, LKB1, and YAP1 Revealed as MYC Interactors with NanoLuc-Based Protein-Fragment Complementation Assay.The c-Myc (MYC) transcription factor is a major cancer driver and a well-validated therapeutic target.  In this study, we report the development of a NanoLuc-based protein-fragment complementation assay (NanoPCA) and mapping of the MYC protein interaction hub in live mammalian cells. 
NanoLuc-based protein-fragment complementation assay
In this study, we report the development of a NanoLuc-based protein-fragment complementation assay (NanoPCA) and mapping of the MYC protein interaction hub in live mammalian cells.  The NanoPCA system was configured to enable detection of protein-protein interactions (PPI) at the endogenous level, as shown with PRAS40 dimerization, and detection of weak interactions, such as PINCH1-NCK2.  Importantly, NanoPCA allows the study of PPI dynamics with reversible interactions.  To demonstrate its utility for large-scale PPI detection in mammalian intracellular environment, we have used NanoPCA to examine MYC interaction with 83 cancer-associated proteins in live cancer cell lines. 
NanoPCA
The NanoPCA system was configured to enable detection of protein-protein interactions (PPI) at the endogenous level, as shown with PRAS40 dimerization, and detection of weak interactions, such as PINCH1-NCK2. 
NanoPCA system
Importantly, NanoPCA allows the study of PPI dynamics with reversible interactions. 
NanoPCA allows the study of PPI dynamics with reversible interactions