title: Acetylcholinesterase immobilized capillary reactors-tandem mass spectrometry: an on-flow tool for ligand screening.
abstract: The use of immobilized capillary enzyme reactors (ICERs) for online ligand screening has been adopted as a new technique for high-throughput screening (HTS). In this work, the selected target was the enzyme acetylcholinesterase (AChE), and the AChE-ICERs produced were used in a liquid chromatograph-tandem ion-trap mass spectrometer. The activity and kinetic parameters were evaluated by monitoring the choline's precursor ion (M + H)(+)m/z 104.0 and its ion fragment (C2H3OH) - (M + H)(+)m/z 60.0. The assay method was validated using the reference AChE inhibitors tacrine and galanthamine. Two new ligands, out of a library of 17 coumarin derivatives, were identified, and the half-maximal inhibitory concentration (IC50), inhibition constant (K(i)), and the inhibition mechanism were determined. A coumarin derivative with IC50 similar to tacrine was highlighted.
authors: Departamento de Química, Universidade Federal de São Carlos, CP 676, São Carlos, 13565-905, SP, Brazil.
authors: Kenia L
authors: KL
authors: Vanzolini
authors: Lucas C C
authors: LC
authors: Vieira
authors: Arlene G
authors: AG
authors: Corrêa
authors: Carmen L
authors: CL
authors: Cardoso
authors: Quezia B
authors: QB
authors: Cass
mesh_terms: D000110:Acetylcholinesterase / Q000378:metabolism*; D002800:Cholinesterase Inhibitors / Q000302:isolation & purification* / Q000494:pharmacology; D002853:Chromatography, Liquid; D003374:Coumarins / Q000302:isolation & purification / Q000494:pharmacology; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005702:Galantamine / Q000494:pharmacology; D057166:High-Throughput Screening Assays / Q000379:methods; D020128:Inhibitory Concentration 50; D007700:Kinetics; D008024:Ligands; D013619:Tacrine / Q000494:pharmacology; D053719:Tandem Mass Spectrometry
chemical_list: D002800:Cholinesterase Inhibitors; D003374:Coumarins; D004800:Enzymes, Immobilized; D008024:Ligands; D005702:Galantamine; D013619:Tacrine; D000110:Acetylcholinesterase