This is an example on how to use the same solubilization methodology to maintain protein-protein interaction and therefore, learn about the partners of a given protein. We could identify the membrane protein partner of BAG3 (Bcl-2 associated athanogene 3). This protein is released by pancreatic ductal adenocarcinoma (PDAC) cells and activates macrophages through a specific unknown receptor.
BAG3 is soluble protein that is known to be present very often on the surface of the macrophage (fluorescence). The idea is to apply our solubilization methodologies to maintain protein-protein interactions and after co-purification, mass spectrometry could identify the protein partner.
One of the partners is IFITM2 (Interferon-induced transmembrane protein 2) has been identified in the experiment but not in the negative control. This was confirmed by other experiments as described here. Complementary Co-IP approach of IFITM2 and by RNAi approach.
We have conducted very similar work for other projects. This is a powerful tool to understand the mechanism of action and identify protein partner in a native environment.
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