title: Surface plasmon resonance based assay for the detection and characterization of promiscuous inhibitors.
abstract: Promiscuous binders achieve enzyme inhibition using a nonspecific aggregation-type binding mechanism to proteins. These compounds are a source of false-positive hits in biochemical inhibition assays and should be removed from screening hit lists because they are not good candidates to initiate medicinal chemistry programs. We introduce a robust approach to identify these molecules early in the lead generation process using real time surface plasmon resonance based biosensors to observe the behavior of the binding interactions between promiscuous compounds and proteins. Furthermore, the time resolution of the assay reveals a number of distinct mechanisms that promiscuous compounds employ to inhibit enzyme function and indicate that the type of mechanism can vary depending on the protein target. A classification scheme for these compounds is presented that can be used to rapidly characterize the hits from high-throughput screens and eliminate compounds with a nonspecific mechanism of inhibition.
authors.affiliation: Roche Palo Alto LLC, Palo Alto, CA 94304, USA. tony.giannetti@roche.com
authors.forename: Anthony M
authors.initials: AM
authors.lastname: Giannetti
authors.forename: Bruce D
authors.initials: BD
authors.lastname: Koch
authors.forename: Michelle F
authors.initials: MF
authors.lastname: Browner
mesh_terms: D015374:Biosensing Techniques; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004791:Enzyme Inhibitors / Q000737:chemistry*; D004798:Enzymes / Q000737:chemistry*; D011485:Protein Binding; D020349:Surface Plasmon Resonance; D013501:Surface-Active Agents
chemical_list: D004791:Enzyme Inhibitors; D004798:Enzymes; D013501:Surface-Active Agents