High-throughput screening of enzyme inhibitors: automatic determination of tight-binding inhibition constants.
Determination of tight-binding inhibition constants by nonlinear least-squares regression requires sufficiently good initial estimates of the best-fit values. Normally an initial estimate of the inhibition constant must be provided by the investigator. This paper describes an automatic procedure for the estimation of tight-binding inhibition constants directly from dose-response data. Because the procedure does not require human intervention, it was incorporated into an algorithm for high-throughput screening of enzyme inhibitors. A suitable computer program is available electronically (http://www.biokin.com). Representative experimental data are shown for the inhibition of human mast-cell tryptase.
Kuzmic
P
P
BioKin, Ltd., Madison, Wisconsin 53708, USA. pkuzmic@biokin.com
Sideris
S
S
Cregar
L M
LM
Elrod
K C
KC
Rice
K D
KD
Janc
J W
JW
D000465:Algorithms; D001331:Automation; D001667:Binding, Competitive; D053818:Chymases; D003198:Computer Simulation; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000494:pharmacology; D006801:Humans; D007700:Kinetics; D009010:Monte Carlo Method; D012697:Serine Endopeptidases / Q000187:drug effects / Q000378:metabolism*; D053802:Tryptases
D004791:Enzyme Inhibitors; D012697:Serine Endopeptidases; C083294:chymase 2; D053818:Chymases; D053802:Tryptases