Project description:Binding partners of CERS6 were identified by co-immunoprecipitation, and a liquid chromatograph and a tandem mass spectrometry analysis
Project description:Co-immunoprecipitation of recombinantly expressed IPO9 using a strep-tag in HeLa cell lysate to identify IPO9 interaction partners.
Project description:The malaria parasite, Plasmodium falciparum, traffics the virulence protein P. falciparum erythrocyte membrane protein 1 (PfEMP1) to the surface of infected red blood cells (RBCs) via membranous organelles known as the Maurer’s clefts. We developed a method for efficient enrichment of Maurer’s clefts and profiled the protein composition of this trafficking organelle. We identified 18 previously uncharacterised or poorly characterised Maurer’s cleft proteins. Transfectants expressing GFP-fusions of 7 of these proteins and confirmed their Maurer’s cleft location. Co-immunoprecipitation and mass spectrometry identified interacting partners of these Maurer’s clefts proteins, providing a network map of protein-protein associations. We identified two key clusters of proteins that may function in the loading and unloading of PfEMP1 into and out of the Maurer’s clefts.
Project description:HA-LUBEL 1-872 or LacZ (control) was expressed under the control of Mef2-GAL4, and the progenies were raised at 25°C. The 3IL carcass fillet lysates were subjected to anti-HA immunoprecipitation. To identify potential binding partners, we performed immunoprecipitation-mass spectrometry (IP-MS) of LUBEL1-872.
Project description:This project contains co-immunoprecipitation (Co-IP) coupled with LC-MS/MS data for the identification of SIRT3-interacting proteins. HEK293T cells were transfected to overexpress SIRT3, followed by immunoprecipitation using an anti-SIRT3 antibody. The immunoprecipitated proteins were separated by SDS-PAGE, subjected to in-gel digestion, and analyzed by mass spectrometry.
Project description:Interacting partners of TSG-6 were analyzed by coimmunoprecipitation (co-IP) coupled with liquid chromatography tandem mass spectrometry (LC–MS/MS)
Project description:We identified GRA57 as a Toxoplasma gondii secreted effector required for parasite survival of interferon gamma in HFFs. To identify interaction partners of GRA57 during infection of HFFs, which could indicate how GRA57 functions to protect the parasite, we performed co-immunoprecipitation (IP) experiments with an endogenously tagged RHGRA57-HA strain.To enable detection of protein-protein interactions specific to activated cells, HFFs were pre-stimulated before infection with 2.5 U/ml IFN for 6 h, which we found was sufficient to induce IFN responses while retaining host cell viability. HFFs were infected for 24 h with either RH∆Ku80 or RHGRA57-HA parasites prior to lysis. GRA57-HA was immunoprecipitated from lysates, then co-immunoprecipitated proteins were identified by liquid chromatography (LC)-tandem mass spectrometry (MS/MS).
Project description:Flag-PFKFB3 was expressed in SW1990 cells. Immunoprecipitation analysis was performed using the anti-Flag antibody, and the extracts were analyzed by mass spectrometry.
Project description:To identify YAP binding partners potentially involved in mechanotransduction, we performed a YAP co-IP/MS experiment using sparsely or densely plated MCF10A cells.