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:To identify the circNEIL3 interacting protein, we performed RNA pull-down assays after transfecting into cells with biotin-labeled sense (S) or antisense (AS) DNA oligomers crossing the junction site of circNEIL3, and the pull-down proteins were subjected to SDS-PAGE and Coomassie blue staining. ~45 kDa Band pulled down by the AS probe was specifically excised , digested and used for mass spectrometry analysis.
Project description:To assess direct involvement of LINC00955 in regulation of gene expression, LINC00955 was used as an RNA probe in RNA pull-down assays and mass spectrometry analyses.