TY - JOUR
T1 - Inhibiting AMPylation
T2 - A novel screen to identify the first small molecule inhibitors of protein AMPylation
AU - Lewallen, Daniel M.
AU - Sreelatha, Anju
AU - Dharmarajan, Venkatasubramanian
AU - Madoux, Franck
AU - Chase, Peter
AU - Griffin, Patrick R.
AU - Orth, Kim
AU - Hodder, Peter
AU - Thompson, Paul R.
PY - 2014/2/21
Y1 - 2014/2/21
N2 - 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 Ki's ranging from 6 to 50 μM and upward of 30-fold selectivity versus HYPE, the only known human AMPylator.
AB - 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 Ki's ranging from 6 to 50 μM and upward of 30-fold selectivity versus HYPE, the only known human AMPylator.
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U2 - 10.1021/cb4006886
DO - 10.1021/cb4006886
M3 - Article
C2 - 24274060
AN - SCOPUS:84896721731
SN - 1554-8929
VL - 9
SP - 433
EP - 442
JO - ACS chemical biology
JF - ACS chemical biology
IS - 2
ER -