Project description:We performed a BioID experiment in a HEK293 Flp-In T-Rex cell line that contains human GNL3 cDNA integrated at the FRT site tagged with BirA and FLAG under the control of a promoter regulated with doxycycline.
Project description:To gain insights into the interactome of wild-type (WT) and S102P mutant GATAD1, we utilized the BioID method, which enables the study of protein-protein interactions. Specifically, we performed BioID proximity labeling experiments in stable Flp-In cells expressing different GATAD1 variants fused to BirA*_FLAG. These variants included BirA*_FLAG_GATAD1-WT, BirA*_FLAG_GATAD1-S102P, BirA*_FLAG_GATAD1-S102D, and BirA*_FLAG_GATAD1-S102A. By employing this approach, we aimed to characterize the protein interactors associated with these GATAD1 variants and gain insights into the functional consequences of the S102P mutation.
Project description:Data contains LC-MS results of proximity-dependent biotin labeling to generate the protein interaction network of IRS1 protein fused with Flag-miniTurbo (N-terminal tag)
and expressed in T-REx HEK293 cells.
Project description:triple SILAC phosphoproteomics experiment of Flp-In T-Rex HEK293 cells stably expressing either FLAG empty, PINK1-FLAG Kinase dead or PINK1-FLAG wild type were grown in ‘light’ (K0R0), ‘medium’ (K4R6) and ‘heavy’ (K8,R10) SILAC media, respectively.
Project description:Affinity purification coupled with mass spectrometry (AP-MS) and proximity-dependent biotinylation identification (BioID) methods are powerful tools to define the interactome for a specific protein bait. Whereas AP-MS results in the identification of proteins that are in a stable complex, BioID labels and identifies proteins that are in close proximity to the bait, resulting in overlapping yet distinct protein identifications. In order to comprehensively characterize the IBTK interactome networks in cells, we developed a tag workflow which allows for both AP-MS and BioID analysis with a single construct, pcDNA5/FRT vector containing FLAG-BirA-IBTK.
Project description:UPF3A and UPF3B are paralogous genes in human cells that are involved in the nonsense-mediated decay (NMD) pathway. NMD is a cellular quality control mechanism that monitors mRNAs during translation. Aberrant translation due to features such as the presence of a premature stop codon downstream on an exon-exon junction activates NMD and leads to the degradation of the mRNA. To investigate the role of UPF3B in NMD, we have generated FLAG-TurboID-UPF3B wild type or mutant expressing human Flp-In T-Rex 293 UPF3B knockout cells using the PiggyBac transposon system. We generated proteomic data from the proximity labeled interactome of these UPF3B variants.