Pan, Sijun Jang, Se-Young Wang, Danyang Liew, Si Si Li, Zhengqiu Lee, Jun-Seok Yao, Shao Q. 2024-01-20T00:32:35Z 2024-01-20T00:32:35Z 2021-09-05 2017-09-18 1433-7851 https://pubs.kist.re.kr/handle/201004/122273 Affinity-based probes (AfBPs) provide a powerful tool for large-scale chemoproteomic studies of drug-target interactions. The development of high-quality probes capable of recapitulating genuine drug-target engagement, however, could be challenging. Minimalist photo-crosslinkers, which contain an alkyl diazirine group and a chemically tractable tag, could alleviate such challenges, but few are currently available. Herein, we have developed new alkyl diazirine-containing photo-crosslinkers with different bioorthogonal tags. They were subsequently used to create a suite of AfBPs based on GW841819X (a small molecule inhibitor of BRD4). Through invitro and insitu studies under conditions that emulated native drug-target interactions, we have obtained better insights into how a tag might affect the probe's performance. Finally, SILAC-based chemoproteomic studies have led to the discovery of a novel off-target, APEX1. Further studies showed GW841819X binds to APEX1 and caused up-regulation of endogenous DNMT1 expression under normoxia conditions. English John Wiley & Sons Ltd. AFFINITY-BASED PROBES TARGET IDENTIFICATION BIOORTHOGONAL LIGATION KINASE INHIBITORS ENGAGEMENT MOLECULES DISCOVERY PROTEINS STRATEGY A Suite of "Minimalist" Photo-Crosslinkers for Live-Cell Imaging and Chemical Proteomics: Case Study with BRD4 Inhibitors Article 10.1002/anie.201706076 1 Angewandte Chemie International Edition, v.56, no.39, pp.11816 - 11821 Angewandte Chemie International Edition 56 39 11816 11821 scie scopus 000410810600024 2-s2.0-85028359967 Chemistry, Multidisciplinary Chemistry Article AFFINITY-BASED PROBES TARGET IDENTIFICATION BIOORTHOGONAL LIGATION KINASE INHIBITORS ENGAGEMENT MOLECULES DISCOVERY PROTEINS STRATEGY affinity-based probes bioorthogonal chemistry epigenetics live-cell imaging photo-crosslinkers