title: Characterization of chemical libraries for luciferase inhibitory activity.
abstract: To aid in the interpretation of high-throughput screening (HTS) results derived from luciferase-based assays, we used quantitative HTS, an approach that defines the concentration-response behavior of each library sample, to profile the ATP-dependent luciferase from Photinus pyralis against more than 70,000 samples. We found that approximately 3% of the library was active, containing only compounds with inhibitory concentration-responses, of which 681 (0.9%) exhibited IC 50 < 10 microM. Representative compounds were shown to inhibit purified P. pyralis as well as several commercial luciferase-based detection reagents but were found to be largely inactive against Renilla reniformis luciferase. Light attenuation by the samples was also examined and found to be more prominent in the blue-shifted bioluminescence produced by R. reniformis luciferase than in the bioluminescence produced by P. pyralis luciferase. We describe the structure-activity relationship of the luciferase inhibitors and discuss the use of this data in the interpretation of HTS results and configuration of luciferase-based assays.
authors: NIH Chemical Genomics Center, National Human Genome Research Institute, National Institutes of Health, Bethesda, Maryland 20892-3370, USA.
authors: Douglas S
authors: DS
authors: Auld
authors: Noel T
authors: NT
authors: Southall
authors: Ajit
authors: A
authors: Jadhav
authors: Ronald L
authors: RL
authors: Johnson
authors: David J
authors: DJ
authors: Diller
authors: Anton
authors: A
authors: Simeonov
authors: Christopher P
authors: CP
authors: Austin
authors: James
authors: J
authors: Inglese
mesh_terms: D000818:Animals; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D048888:Fireflies / Q000201:enzymology; D008156:Luciferases / Q000037:antagonists & inhibitors*; D008958:Models, Molecular; D013329:Structure-Activity Relationship
chemical_list: D004791:Enzyme Inhibitors; D008156:Luciferases