Equations of substrate-inhibition kinetics applied to pig kidney diamine oxidase (DAO, E.C. 1.4.3.6).
Pig kidney diamine oxidase (DAO) and other semicarbazide-sensitive amine oxidases (SSAO) show clear substrate-inhibition kinetics and a reaction-scheme mechanism based on two substrate binding sites. We evaluated several reaction scheme mechanisms with a non-linear regression program (NCSS), estimating R2, the constants of the equations and their standard errors and we determined the deviation of experimental data from theoretical equations. The best fit was obtained with a "dead end" mechanism with two binding sites. Based on this scheme, other schemes for a two-substrate reaction and for mechanisms of inhibition were constructed. These reaction schemes, even at low substrate concentration, fitted experimental data better than Michaelis-Menten kinetics, and provided information on the mechanisms of action of inhibitors. The presence of two substrate-binding sites on pig kidney DAO was confirmed by all experimental data.
Ignesti
G
G
Department of Pharmacology, University of Florence, V.le G. Pieraccini 6, 50139 Firenze, Italy. ignesti@pharm.unifi.it
D006631:Amine Oxidase (Copper-Containing) / Q000037:antagonists & inhibitors* / Q000378:metabolism; D000818:Animals; D001596:Benzylamines / Q000494:pharmacology; D001665:Binding Sites; D002103:Cadaverine / Q000494:pharmacology; D003959:Diamines / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006632:Histamine / Q000494:pharmacology; D007668:Kidney / Q000187:drug effects / Q000201:enzymology*; D007700:Kinetics; D008956:Models, Chemical; D011700:Putrescine / Q000494:pharmacology; D051381:Rats; D013095:Spermidine / Q000494:pharmacology; D013379:Substrate Specificity; D013552:Swine
D001596:Benzylamines; D003959:Diamines; D004791:Enzyme Inhibitors; C015127:1,7-diaminoheptane; D006632:Histamine; C030796:benzylamine; D006631:Amine Oxidase (Copper-Containing); D002103:Cadaverine; D013095:Spermidine; D011700:Putrescine; C007297:1,6-diaminohexane