Development of a novel helicase assay using electrochemiluminescence.
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.
Zhang
L
L
Lead Discovery, Aventis Pharmaceuticals, 500 Arcola Road, NW15, Collegeville, PA 19426, USA. Litao.zhang@dupontpharma.com
Schwartz
G
G
O'Donnell
M
M
Harrison
R K
RK
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
D017931:DNA Primers; D004265:DNA Helicases