title: Lathyrus cicera copper amine oxidase reactions with tryptamine.
abstract: Lathyrus cicera copper amine oxidase (LCAO) rapidly formed the typical Cu(I)-TPQ semiquinone UV-visible spectrum, identical to that formed by other substrates, upon O(2) exhaustion by turnover with excess tryptamine. A new band at 630 nm formed more slowly, with intensity dependent on aldehyde and H(2)O(2) concentrations. On prolonged incubation, all bands decayed in parallel, together with loss of enzymatic activity. The blue color disappeared on addition of KCN, a Cu(I) stabilizing agent, while the intensity of the radical visible bands increased. This shows that the 630 nm absorbing species is a Cu(II) derivative, as confirmed by the unchanged intensity of the EPR spectrum of the frozen blue solution from that of the native protein. Rapid kinetics experiments showed that this species derives from a reduced form of the protein, plus aldehyde and H(2)O(2) and that it is not in dynamic equilibrium with the radical. Given the similar population of the semiquinone radical with all substrates, it is possible that the reaction with aldehyde and H(2)O(2) occurs in all cases although substrates lacking the indole group only produce the Cu(I)-semiquinone band. The radical participation to the catalytic activity is demonstrated by the observation that its relative population (controlled by the pH) parallels changes in the reoxidation rate constant, while the 630 nm absorbing species is implied in the inactivation process, which depends on H(2)O(2) and aldehyde concentration. The results of the paper are consistent with half-of-the-site reactivity, i.e. the two subunits of LCAO are kinetically and spectroscopically distinct from each other.
authors.affiliation: Department of Biochemical Sciences, A. Rossi Fanelli, University of Rome Sapienza, P.le A. Moro 5, 00185 Rome, Italy. paola.pietrangeli@uniroma1.it
authors.forename: Paola
authors.initials: P
authors.lastname: Pietrangeli
authors.forename: Andrea
authors.initials: A
authors.lastname: Bellelli
authors.forename: Paola
authors.initials: P
authors.lastname: Fattibene
authors.forename: Bruno
authors.initials: B
authors.lastname: Mondovì
authors.forename: Laura
authors.initials: L
authors.lastname: Morpurgo
mesh_terms: D006631:Amine Oxidase (Copper-Containing) / Q000378:metabolism*; D004578:Electron Spin Resonance Spectroscopy; D007700:Kinetics; D027807:Lathyrus / Q000201:enzymology*; D012701:Serotonin / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D014363:Tryptamines / Q000378:metabolism*
chemical_list: D014363:Tryptamines; D012701:Serotonin; D006631:Amine Oxidase (Copper-Containing)