title: A high-throughput screen utilizing the fluorescence of riboflavin for identification of lumazine synthase inhibitors.
abstract: A high-throughput screening method based on the competitive binding of a lumazine synthase inhibitor and riboflavin to the active site of Schizosaccharomyces pombe lumazine synthase was developed. This assay is sensitive, simple, and robust. During assay development, all of the known active inhibitors tested were positively identified. Preliminary high-throughput screening in 384-well format resulted in a Z factor of 0.7. The approach utilizes a thermodynamic assay to bypass the problems associated with the instabilities of both lumazine synthase substrates that complicate the use of a kinetic assay in a high-throughput format, and it removes the time element from the assay, thus simplifying the procedure.
authors.affiliation: Department of Medicinal Chemistry and Molecular Pharmacology, School of Pharmacy and Pharmaceutical Sciences, Purdue University, West Lafayette, IN 47907, USA.
authors.forename: Jinhua
authors.initials: J
authors.lastname: Chen
authors.forename: Boris
authors.initials: B
authors.lastname: Illarionov
authors.forename: Adelbert
authors.initials: A
authors.lastname: Bacher
authors.forename: Markus
authors.initials: M
authors.lastname: Fischer
authors.forename: Ilka
authors.initials: I
authors.lastname: Haase
authors.forename: Gunda
authors.initials: G
authors.lastname: Georg
authors.forename: Qi-Zhuang
authors.initials: QZ
authors.lastname: Ye
authors.forename: Zeqiang
authors.initials: Z
authors.lastname: Ma
authors.forename: Mark
authors.initials: M
authors.lastname: Cushman
mesh_terms: D001665:Binding Sites; D004353:Drug Evaluation, Preclinical; D004791:Enzyme Inhibitors / Q000302:isolation & purification*; D008024:Ligands; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors*; D012256:Riboflavin / Q000737:chemistry*; D012568:Schizosaccharomyces / Q000201:enzymology; D013050:Spectrometry, Fluorescence
chemical_list: D004791:Enzyme Inhibitors; D008024:Ligands; D009097:Multienzyme Complexes; C044822:6,7-dimethyl-8-ribityllumazine synthase; D012256:Riboflavin