title: On the purification and preliminary crystallographic analysis of isoquinoline 1-oxidoreductase from Brevundimonas diminuta 7.
abstract: Isoquinoline 1-oxidoreductase (IOR) from Brevundimonas diminuta is a mononuclear molybdoenzyme of the xanthine-dehydrogenase family of proteins and catalyzes the conversion of isoquinoline to isoquinoline-1-one. Its primary sequence and behaviour, specifically in its substrate specificity and lipophilicity, differ from other members of the family. A crystal structure of the enzyme is expected to provide an explanation for these differences. This paper describes the crystallization and preliminary X-ray diffraction experiments as well as an optimized purification protocol for IOR. Crystallization of IOR was achieved using two different crystallization buffers. Streak-seeding and cross-linking were essential to obtain well diffracting crystals. Suitable cryo-conditions were found and a structure solution was obtained by molecular replacement. However, phases need to be improved in order to obtain a more interpretable electron-density map.
authors.affiliation: REQUIMTE/CQFB, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal.
authors.forename: D Roeland
authors.initials: DR
authors.lastname: Boer
authors.forename: Axel
authors.initials: A
authors.lastname: Müller
authors.forename: Susanne
authors.initials: S
authors.lastname: Fetzner
authors.forename: David J
authors.initials: DJ
authors.lastname: Lowe
authors.forename: Maria João
authors.initials: MJ
authors.lastname: Romão
mesh_terms: D002384:Catalysis; D042063:Caulobacteraceae / Q000201:enzymology*; D003460:Crystallization; D019281:Dimerization; D008958:Models, Molecular; D044925:Oxidoreductases Acting on CH-CH Group Donors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D017433:Protein Structure, Secondary; D014961:X-Ray Diffraction
chemical_list: D044925:Oxidoreductases Acting on CH-CH Group Donors; C086842:isoquinoline 1-oxidoreductase