1. A Suite of 'Minimalist' Photo-Crosslinkers for Live-Cell Imaging and Chemical Proteomics: Case Study with BRD4 Inhibitors.
- Author
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Pan, Sijun, Jang, Se‐Young, Wang, Danyang, Liew, Si Si, Li, Zhengqiu, Lee, Jun‐Seok, and Yao, Shao Q.
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PHOTOCROSSLINKING , *CELL imaging , *PROTEOMICS , *TARGETED drug delivery , *ENZYME inhibitors , *SMALL molecules - Abstract
Affinity-based probes (A fBPs) 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 A fBPs based on GW841819X (a small molecule inhibitor of BRD4). Through in vitro and in situ 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. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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