title: Effect of metal substitution in copper amine oxidase from lentil seedlings.
abstract: The reaction with substrates and carbonyl reagents of native lentil Cu-amine oxidase and its modified forms, i.e. Cu-fully-depleted, Cu-half-reconstituted, Cu-fully-reconstituted, Co-substituted, Ni-substituted and Zn-substituted, has been studied. Upon removal of only one of the two Cu ions, the enzyme loses 50% of its enzymatic activity. Using several substrates, Co-substituted lentil amine oxidase is shown to be active but the k(c) value is different from that of native or Cu-fully-reconstituted enzyme, while K(m) is similar. On the other hand, the Ni- and Zn-substituted forms are catalytically inactive. Enzymatic activity measurements and optical spectroscopy show that only in the Co-substituted enzyme is the organic cofactor 6-hydroxydopa quinone reactive and the enzyme catalytically competent, although less efficient. The Co-substituted amine oxidase does not form the semiquinone radical as an intermediate of the catalytic reaction. While devoid or reduced of catalytic activity, all the enzyme preparations are still able to oxidise two moles of substrate and to release two moles of aldehyde per mole of dimeric enzyme. The results obtained show that although Co-substituted amine oxidase is catalytically competent, copper is essential for the catalytic mechanism.
authors.affiliation: Department of Sciences Applied to Biosystems University of Cagliari, I-09042 Cagliari, Italy. padiglia@unica.it
authors.forename: A
authors.initials: A
authors.lastname: Padiglia
authors.forename: R
authors.initials: R
authors.lastname: Medda
authors.forename: J Z
authors.initials: JZ
authors.lastname: Pedersen
authors.forename: A
authors.initials: A
authors.lastname: Finazzi Agrò
authors.forename: A
authors.initials: A
authors.lastname: Lorrai
authors.forename: B
authors.initials: B
authors.lastname: Murgia
authors.forename: G
authors.initials: G
authors.lastname: Floris
mesh_terms: D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry* / Q000378:metabolism; D001596:Benzylamines / Q000378:metabolism; D003300:Copper / Q000737:chemistry; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives / Q000737:chemistry; D007887:Fabaceae / Q000201:enzymology*; D006834:Hydrazines / Q000378:metabolism; D007735:Kynuramine / Q000378:metabolism; D008670:Metals; D010084:Oxidation-Reduction; D010944:Plants / Q000201:enzymology; D010946:Plants, Medicinal; D013379:Substrate Specificity; D015032:Zinc / Q000737:chemistry
chemical_list: D001596:Benzylamines; D006834:Hydrazines; D008670:Metals; C001388:6-hydroxydopa; D007735:Kynuramine; D004295:Dihydroxyphenylalanine; D003300:Copper; C030796:benzylamine; D006631:Amine Oxidase (Copper-Containing); D015032:Zinc