Technical Platform
Chemoproteomic has advanced drug target discovery, especially with chemical probe-based platforms. However, challenges like complex synthesis, low yield, the high cost, and unknown SAR of drug molecules hinder progress. The photoaffinity magnetic bead platform addresses these issues by immobilizing active drug molecules onto functional magnetic beads via UV illumination,eliminating the need for chemical probe design. This ensures specific binding between the drug and its target protein. After incubation with the proteome,the beads enrich and isolate drug target proteins,which are then analyzed by immunoblotting and mass spectrometry.
Platform Features
Case Study
To validate the targets of staurosporine (Stau), we immobilized it onto the surface of photoaffinity-modified functional magnetic beads (Stau-beads). Subsequently, we incubated the Stau-beads with the whole proteome, using Fmoc-beads as a control group. The resulting Western blot (WB) results are shown in the figure below, indicating that Stau-beads effectively enriched the target protein FECH (a known positive target).
Next, we subjected the proteins enriched and separated using Fmoc-beads and Stau-beads to enzymatic digestion and mass spectrometry analysis. We quantitatively compared the intensity differences of proteins between the experimental group (Stau-beads) and the control group (Fmoc-beads). The results demonstrated that all known positive target proteins were significantly enriched in the Stau-beads group, confirming the effectiveness of the method.
