title: Development of a novel helicase assay using electrochemiluminescence.
abstract: DNA helicases are ubiquitous enzymes involved in DNA replication, recombination, and repair. These enzymes use the energy of ATP to unwind duplex DNA. A common approach to measure DNA helicase activity utilizes electrophoresis to visualize product formation in a gel. The present study develops a rapid helicase assay based on electrochemiluminescence. The assay is adaptable to rapid high-throughput screening for chemical inhibitors. The assay is applied to Escherichia coli DnaB. Three other technologies have also been applied to DnaB and are compared to results of the electrochemiluminescence-based assay.
authors.affiliation: Lead Discovery, Aventis Pharmaceuticals, 500 Arcola Road, NW15, Collegeville, PA 19426, USA. Litao.zhang@dupontpharma.com
authors.forename: L
authors.initials: L
authors.lastname: Zhang
authors.forename: G
authors.initials: G
authors.lastname: Schwartz
authors.forename: M
authors.initials: M
authors.lastname: O'Donnell
authors.forename: R K
authors.initials: RK
authors.lastname: Harrison
mesh_terms: D001483:Base Sequence; D004265:DNA Helicases / Q000032:analysis* / Q000378:metabolism; D017931:DNA Primers; D004926:Escherichia coli / Q000201:enzymology*; D005453:Fluorescence; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D012588:Scintillation Counting; D013379:Substrate Specificity
chemical_list: D017931:DNA Primers; D004265:DNA Helicases