Project description:Diaminoquinazolines represent a privileged scaffold for antimalarial discovery, including use as putative Plasmodium histone lysine methyltransferase inhibitors (BIX-01294). Despite this, robust evidence for their molecular targets, proteome-wide, is lacking. Here we report the design and development of a small-molecule photo-crosslinkable probe to investigate the targets of our diaminoquinazoline series. We demonstrate the effectiveness of our designed probe for photoaffinity labelling of Plasmodium lysates and initial pull-down proteomics experiments identified proteins from different classes enriched by the probe, highlighting the suitability of the developed probe as a valuable tool for protein identification in Plasmodium falciparum.
Project description:Given the pivotal role of Tat in HIV-1 latency, we performed RNA pull-down assays in latently HIV-1-infected J-Lat 10.6 cells using biotinylated RNA probes (including a control RNA probe and a Tat-specific RNA probe) . Cell lysates were incubated with streptavidin agarose resin pre-bound to Tat RNA probes, and the enriched Tat RNA-binding proteins were separated by SDS-PAGE and analyzed by liquid chromatography mass spectrometry (LC-MS/MS)
Project description:Circular RNAs (circRNAs) have emerged as critical regulators in various biological processes through their interactions with RNA-binding proteins. To investigate the protein interactions of hsa_circ_0001910, a circRNA of potential functional significance, we designed a specific RNA pulldown probe targeting this molecule. Using this probe, we performed RNA pull-down assays to enrich proteins that directly or indirectly bind to hsa_circ_0001910. The pulled-down protein complexes were subsequently separated and detected by Western blotting to confirm the presence of specific protein candidates. For comprehensive and unbiased protein identification, the enriched protein samples were further subjected to mass spectrometry (MS)-based proteomic analysis. This approach allows for the systematic identification and quantification of proteins that interact with hsa_circ_0001910, providing insights into its molecular functions and potential roles in cellular pathways.