title: Purification and characterization of pea seedling amine oxidase for crystallization studies.
abstract: Pea (Pisum sativum L.) seedling amine oxidase (EC 1.4.3.6) is the first amine oxidase to be crystallized that diffracts to atomic resolution (2.5 A). Extensive modifications of a published purification procedure were necessary to obtain protein that would give diffraction-quality crystals. Here we report the improved purification and also use this high-purity protein to reexamine some fundamental characteristics of pea seedling amine oxidase. The extinction coefficient at 280 nm (epsilon 1%(280)) and the molecular mass of the protein are investigated by a variety of techniques, yielding epsilon 1%(280) = 20 cm-1 and a mass 150 +/- 6 kD. In addition, the stoichiometry of the metal and organic cofactors, Cu(II) and 6-hydroxy dopa (Topa) quinone, respectively, is examined. The ratio of Cu(II):Topa:protein monomer is found to be 1:1:1.
authors: Department of Chemistry and Biochemistry, Montana State University, Bozeman 59717.
authors: M A
authors: MA
authors: McGuirl
authors: C D
authors: CD
authors: McCahon
authors: K A
authors: KA
authors: McKeown
authors: D M
authors: DM
authors: Dooley
mesh_terms: D006631:Amine Oxidase (Copper-Containing); D002845:Chromatography; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003300:Copper / Q000032:analysis; D018360:Crystallography, X-Ray; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives / Q000032:analysis; D017886:Durapatite; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae / Q000201:enzymology*; D008970:Molecular Weight; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000737:chemistry* / Q000302:isolation & purification*; D010946:Plants, Medicinal; D013053:Spectrophotometry
chemical_list: D004295:Dihydroxyphenylalanine; C067626:6-hydroxydopa quinone; D003300:Copper; D017886:Durapatite; D006631:Amine Oxidase (Copper-Containing); D000587:Oxidoreductases Acting on CH-NH Group Donors