title: High-resolution crystal structure of copper amine oxidase from Arthrobacter globiformis: assignment of bound diatomic molecules as O2.
abstract: The crystal structure of a copper amine oxidase from Arthrobacter globiformis was determined at 1.08 Å resolution with the use of low-molecular-weight polyethylene glycol (LMW PEG; average molecular weight ∼200) as a cryoprotectant. The final crystallographic R factor and Rfree were 13.0 and 15.0%, respectively. Several molecules of LMW PEG were found to occupy cavities in the protein interior, including the active site, which resulted in a marked reduction in the overall B factor and consequently led to a subatomic resolution structure for a relatively large protein with a monomer molecular weight of ∼70,000. About 40% of the presumed H atoms were observed as clear electron densities in the Fo - Fc difference map. Multiple minor conformers were also identified for many residues. Anisotropic displacement fluctuations were evaluated in the active site, which contains a post-translationally derived quinone cofactor and a Cu atom. Furthermore, diatomic molecules, most likely to be molecular oxygen, are bound to the protein, one of which is located in a region that had previously been proposed as an entry route for the dioxygen substrate from the central cavity of the dimer interface to the active site.
authors: Department of Biochemistry, Osaka Medical College, 2-7 Daigakumachi, Takatsuki, Osaka 569-8686, Japan.
authors: Takeshi
authors: T
authors: Murakawa
authors: Hideyuki
authors: H
authors: Hayashi
authors: Tomoko
authors: T
authors: Sunami
authors: Kazuo
authors: K
authors: Kurihara
authors: Taro
authors: T
authors: Tamada
authors: Ryota
authors: R
authors: Kuroki
authors: Mamoru
authors: M
authors: Suzuki
authors: Katsuyuki
authors: K
authors: Tanizawa
authors: Toshihide
authors: T
authors: Okajima
mesh_terms: D006631:Amine Oxidase (Copper-Containing) / Q000737:chemistry*; D016880:Anisotropy; D001173:Arthrobacter / Q000737:chemistry / Q000201:enzymology*; D001665:Binding Sites; D003451:Cryoprotective Agents / Q000737:chemistry; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D010100:Oxygen / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011487:Protein Conformation
chemical_list: D003451:Cryoprotective Agents; D011092:Polyethylene Glycols; D006631:Amine Oxidase (Copper-Containing); D010100:Oxygen
keywords: TPQ; copper amine oxidase