caa a22 4500 445376481 CHVBK 20180317143004.0 cr unu---uuuuu 170323e20110701xx s 000 0 eng 10.1007/s00702-011-0657-y doi (NATIONALLICENCE)springer-10.1007/s00702-011-0657-y An improved approach to steady-state analysis of monoamine oxidases [Elektronische Daten] [Rona Ramsay, Aldo Olivieri, Andrew Holt] The search for new monoamine oxidase inhibitors aims to identify potential lead compounds that are more potent and selective than current drugs for use in treating a variety of neuropsychiatric and neurodegenerative conditions. An integral part of this process is a kinetic examination of monoamine oxidases in the presence of the inhibitor, to determine potency and selectivity and to obtain information on mechanism. To date, kinetic data obtained with a probe substrate have been analysed by fitting to the Michaelis-Menten equation which describes a unireactant process in which velocity is related to substrate concentration in a rectangular hyperbolic manner. In this study, we present evidence that monoamine oxidase activity is often not adequately described by this approach. We outline a novel equation strategy that takes account of substrate and inhibitor binding to oxidised and reduced enzyme forms, and quantifies differences between substrates and inhibitors in this regard. When combined with plate reader-based experimental techniques that allow large numbers of substrate and inhibitor concentrations to be used, and the global nonlinear regression facilities of GraphPad Prism software, this straightforward approach allows more appropriate analyses of monoamine oxidase by non-experts than has previously been possible. Springer-Verlag, 2011 Monoamine oxidase nationallicence Steady-state kinetics nationallicence Equations nationallicence Inhibition nationallicence GraphPad Prism nationallicence Ramsay Rona School of Biology, Biomedical Sciences Research Complex, University of St. Andrews, BMS Building, North Haugh, St. Andrews, KY16 9ST, Fife, UK aut Olivieri Aldo Department of Pharmacology, Faculty of Medicine and Dentistry, University of Alberta, 9-70 Medical Sciences Building, T6G 2H7, Edmonton, AB, Canada aut Holt Andrew Department of Pharmacology, Faculty of Medicine and Dentistry, University of Alberta, 9-70 Medical Sciences Building, T6G 2H7, Edmonton, AB, Canada aut Journal of Neural Transmission Springer Vienna 118/7(2011-07-01), 1003-1019 0300-9564 118:7<1003 2011 118 702 https://doi.org/10.1007/s00702-011-0657-y text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00702-011-0657-y text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ramsay Rona School of Biology, Biomedical Sciences Research Complex, University of St. Andrews, BMS Building, North Haugh, St. Andrews, KY16 9ST, Fife, UK aut NATIONALLICENCE 700 1- Olivieri Aldo Department of Pharmacology, Faculty of Medicine and Dentistry, University of Alberta, 9-70 Medical Sciences Building, T6G 2H7, Edmonton, AB, Canada aut NATIONALLICENCE 700 1- Holt Andrew Department of Pharmacology, Faculty of Medicine and Dentistry, University of Alberta, 9-70 Medical Sciences Building, T6G 2H7, Edmonton, AB, Canada aut NATIONALLICENCE 773 0- Journal of Neural Transmission Springer Vienna 118/7(2011-07-01), 1003-1019 0300-9564 118:7<1003 2011 118 702 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer