Inhibiting AMPylation: A Novel Screen To Identify the First Small Molecule Inhibitors of Protein AMPylation

详细信息    查看全文

• 作者：Daniel M. Lewallen ; Anju Sreelatha ; Venkatasubramanian Dharmarajan ; Franck Madoux ; Peter Chase ; Patrick R. Griffin ; Kim Orth ; Peter Hodder ; Paul R. Thompson
• 刊名：ACS Chemical Biology
• 出版年：2014
• 出版时间：February 21, 2014
• 年：2014
• 卷：9
• 期：2
• 页码：433-442
• 全文大小：624K
• ISSN：1554-8937

文摘

Enzymatic transfer of the AMP portion of ATP to substrate proteins has recently been described as an essential mechanism of bacterial infection for several pathogens. The first AMPylator to be discovered, VopS from Vibrio parahemolyticus, catalyzes the transfer of AMP onto the host GTPases Cdc42 and Rac1. Modification of these proteins disrupts downstream signaling events, contributing to cell rounding and apoptosis, and recent studies have suggested that blocking AMPylation may be an effective route to stop infection. To date, however, no small molecule inhibitors have been discovered for any of the AMPylators. Therefore, we developed a fluorescence-polarization-based high-throughput screening assay and used it to discover the first inhibitors of protein AMPylation. Herein we report the discovery of the first small molecule VopS inhibitors (e.g., calmidazolium, GW7647, and MK886) with K_i鈥檚 ranging from 6 to 50 渭M and upward of 30-fold selectivity versus HYPE, the only known human AMPylator.