title: An ultraefficient affinity-based high-throughout screening process: application to bacterial cell wall biosynthesis enzyme MurF.
abstract: The authors describe the discovery of a new class of inhibitors to an essential Streptococcus pneumoniae cell wall biosyn-thesis enzyme, MurF, by a novel affinity screening method. The strategy involved screening very large mixtures of diverse small organic molecules against the protein target on the basis of equilibrium binding, followed by iterative ultrafiltration steps and ligand identification by mass spectrometry. Hits from any affinity-based screening method often can be relatively nonselective ligands, sometimes referred to as "nuisance" or "promiscuous" compounds. Ligands selective in their binding affinity for the MurF target were readily identified through electronic subtraction of an empirically determined subset of promiscuous compounds in the library without subsequent selectivity panels. The complete strategy for discovery and identification of novel specific ligands can be applied to all soluble protein targets and a wide variety of ligand libraries.
authors.affiliation: Department of Target and Lead Discovery, Global Pharmaceutical R&D, Abbott Laboratories, Abbott Park, Illinois 60064-6217, USA. kenneth.m.comess@abbott.com
authors.forename: Kenneth M
authors.initials: KM
authors.lastname: Comess
authors.forename: Mark E
authors.initials: ME
authors.lastname: Schurdak
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Voorbach
authors.forename: Michael
authors.initials: M
authors.lastname: Coen
authors.forename: Jonathan D
authors.initials: JD
authors.lastname: Trumbull
authors.forename: Houjun
authors.initials: H
authors.lastname: Yang
authors.forename: Lan
authors.initials: L
authors.lastname: Gao
authors.forename: Hua
authors.initials: H
authors.lastname: Tang
authors.forename: Xueheng
authors.initials: X
authors.lastname: Cheng
authors.forename: Claude G
authors.initials: CG
authors.lastname: Lerner
authors.forename: J Owen
authors.initials: JO
authors.lastname: McCall
authors.forename: David J
authors.initials: DJ
authors.lastname: Burns
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Beutel
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology*; D002473:Cell Wall / Q000378:metabolism*; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D013058:Mass Spectrometry; D008826:Microbial Sensitivity Tests; D010453:Peptide Synthases / Q000037:antagonists & inhibitors*; D013296:Streptococcus pneumoniae / Q000201:enzymology*
chemical_list: D000900:Anti-Bacterial Agents; D010453:Peptide Synthases; C060628:UDP-N-acetylmuramoylalanyl-D-glutamyllysine-D-alanyl-D-alanine ligase