Purification and Characterization of Pea Seedling Amine Oxidase for Crystallization Studies
Open Access
- 1 November 1994
- journal article
- Published by Oxford University Press (OUP) in Plant Physiology
- Vol. 106 (3) , 1205-1211
- https://doi.org/10.1104/pp.106.3.1205
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 of 150 [plus or minus] 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.Keywords
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