Kynuramine, a fluorescent substrate and probe of plasma amine oxidase

The fluorescence substrate kynuramine was used as a probe of the catalytic site of plasma amine oxidase. Under anaerobic conditions, the binding of kynuramine causes several spectroscopic changes. The Stokes shift (deltav = 5326 cm-) associated with binding of the substrate to the Enzyme can be attributed to nonpolar properties of the binding site, whereas the increase in emission anisotropy (A = 33) indicates rigid attachment of the substrate to the Enzyme. The fluorescence enhancement that follows the binding of substrate was used to determine the association constant (Ka). The Enzyme plasma amine oxidase binds only 1 molecule of substrate with a Ka = 1.8 X 10(5) M-1 under anaerobic conditions. The use of fluorescence substrates seems to offer the possibility of monitoring conformational changes occurring prior to the catalytic event.