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Harnessing the anti-cancer natural product nimbolide for targeted protein degradation.

Nat. Chem. Biol.. 2019-07; 
SpradlinJessica N,HuXirui,WardCarl C,BrittainScott M,JonesMichael D,OuLisha,ToMilton,ProudfootAndrew,OrnelasElizabeth,WoldegiorgisMikias,OlzmannJames A,BussiereDirksen E,ThomasJason R,TallaricoJohn A,McKennaJeffrey M,SchirleMarkus,MaimoneThomas J,NomuraDani
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Peptide Synthesis After 48 h cells were collected in TBS, lysed by sonication and batch bound with anti-DYKDDDDK resin (GenScript, L00432) for 90 min. Lysate and resin was loaded onto a gravity flow column and washed, followed by elution with 250 ng μl −1 3XFLAG peptide (ApexBio, A6001). Get A Quote

摘要

Nimbolide, a terpenoid natural product derived from the Neem tree, impairs cancer pathogenicity; however, the direct targets and mechanisms by which nimbolide exerts its effects are poorly understood. Here, we used activity-based protein profiling (ABPP) chemoproteomic platforms to discover that nimbolide reacts with a novel functional cysteine crucial for substrate recognition in the E3 ubiquitin ligase RNF114. Nimbolide impairs breast cancer cell proliferation in-part by disrupting RNF114-substrate recognition, leading to inhibition of ubiquitination and degradation of tumor suppressors such as p21, resulting in their rapid stabilization. We further demonstrate that nimbolide can be harnessed to r... More

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