A FluoPol-ABPP PAD2 High-Throughput Screen Identifies the First Calcium Site Inhibitor Targeting the PADs

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• 作者：Daniel M. Lewallen ; Kevin L. Bicker ; Franck Madoux ; Peter Chase ; Lynne Anguish ; Scott Coonrod ; Peter Hodder ; Paul R. Thompson
• 刊名：ACS Chemical Biology
• 出版年：2014
• 出版时间：April 18, 2014
• 年：2014
• 卷：9
• 期：4
• 页码：913-921
• 全文大小：463K
• 年卷期：v.9,no.4(April 18, 2014)
• ISSN：1554-8937

文摘

The protein arginine deiminases (PADs) catalyze the post-translational hydrolysis of peptidyl-arginine to form peptidyl-citrulline in a process termed deimination or citrullination. PADs likely play a role in the progression of a range of disease states because dysregulated PAD activity is observed in a host of inflammatory diseases and cancer. For example, recent studies have shown that PAD2 activates ER伪 target gene expression in breast cancer cells by citrullinating histone H3 at ER target promoters. To date, all known PAD inhibitors bind directly to the enzyme active site. PADs, however, also require calcium ions to drive a conformational change between the inactive apo-state and the fully active calcium bound holoenzyme, suggesting that it would be possible to identify inhibitors that bind the apoenzyme and prevent this conformational change. As such, we set out to develop a screen that can identify PAD2 inhibitors that bind to either the apo or calcium bound form of PAD2. Herein, we provide definitive proof of concept for this approach and report the first PAD inhibitor, ruthenium red (K_i of 17 渭M), to preferentially bind the apoenzyme.