title: High-throughput, cell-based screens to identify small-molecule inhibitors of ricin toxin and related category b ribosome inactivating proteins (RIPs).
abstract: Ricin is a member of the ubiquitous ribosome-inactivating protein (RIP) family of toxins. The Centers for Disease Control and Prevention (CDC) classify ricin and related toxins as Category B biothreat agents. There are currently no antidotes or therapeutics to counteract RIPs in humans. The discovery of effective small-molecule inhibitors of RIPs is increasingly possible, however, due to the availability and accessibility of diverse small-molecule chemical libraries coupled with robust robotics and automated screening methodologies. In this article, we describe a cell-based, high-throughput screening strategy and secondary assays that we have successfully used to identify compounds that target ricin toxin's enzymatic activity and intracellular trafficking, as well as stress-activated signaling pathways associated with cell death. The methods described in the protocol are amenable to the other RIPs.
authors: Division of Infectious Diseases, Wadsworth Center, New York State Department of Health, Albany, New York, USA.
authors: Paul G
authors: PG
authors: Wahome
authors: Nicholas J
authors: NJ
authors: Mantis
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002619:Chemical Warfare Agents; D002522:Chlorocebus aethiops; D057166:High-Throughput Screening Assays / Q000379:methods*; D054788:Ribosome Inactivating Proteins / Q000037:antagonists & inhibitors; D012276:Ricin / Q000037:antagonists & inhibitors*; D054852:Small Molecule Libraries / Q000494:pharmacology*; D014709:Vero Cells
chemical_list: D002619:Chemical Warfare Agents; D054852:Small Molecule Libraries; D012276:Ricin; D054788:Ribosome Inactivating Proteins