title: Inhibiting AMPylation: a novel screen to identify the first small molecule inhibitors of protein AMPylation.
abstract: 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.
authors.affiliation: Department of Chemistry, The Scripps Research Institute, Scripps Florida , 120 Scripps Way, Jupiter, Florida 33458, United States.
authors.forename: Daniel M
authors.initials: DM
authors.lastname: Lewallen
authors.forename: Anju
authors.initials: A
authors.lastname: Sreelatha
authors.forename: Venkatasubramanian
authors.initials: V
authors.lastname: Dharmarajan
authors.forename: Franck
authors.initials: F
authors.lastname: Madoux
authors.forename: Peter
authors.initials: P
authors.lastname: Chase
authors.forename: Patrick R
authors.initials: PR
authors.lastname: Griffin
authors.forename: Kim
authors.initials: K
authors.lastname: Orth
authors.forename: Peter
authors.initials: P
authors.lastname: Hodder
authors.forename: Paul R
authors.initials: PR
authors.lastname: Thompson
mesh_terms: D000249:Adenosine Monophosphate / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D055808:Drug Discovery; D057166:High-Throughput Screening Assays; D006801:Humans; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*; D014735:Vibrio Infections / Q000188:drug therapy / Q000382:microbiology; D014736:Vibrio parahaemolyticus / Q000187:drug effects* / Q000378:metabolism*
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D054852:Small Molecule Libraries; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate