title: Expression, purification, crystallization and preliminary X-ray analysis of sepiapterin reductase from Chlorobium tepidum.
abstract: Sepiapterin reductase from Chlorobium tepidum (CT-SR) produces L-threo-tetrahydrobiopterin, an isomer of tetrahydrobiopterin, in the last step of de novo synthesis initiating from GTP. Native CT-SR and a selenomethionine (SeMet) derivative of CT-SR have been crystallized by the hanging-drop vapour-diffusion method using PEG 400 as precipitant. CT-SR crystals belong to space group R32, with unit-cell parameters a = b = 201.142, c = 210.184 A, and contain four molecules in the asymmetric unit. Diffraction data were collected to 2.1 A resolution using synchrotron radiation. The structure of CT-SR has been determined using MAD phasing. There is one CT-SR tetramer in the asymmetric unit formed by two closely interacting CT-SR dimers. The solvent content is calculated to be about 67.2%.
authors.affiliation: Division of Applied Life Science, Gyeongsang National University, Jinju 660-711, South Korea.
authors.forename: Supangat
authors.initials: S
authors.lastname: Supangat
authors.forename: Yong Kee
authors.initials: YK
authors.lastname: Choi
authors.forename: Young Shik
authors.initials: YS
authors.lastname: Park
authors.forename: Daeyoung
authors.initials: D
authors.lastname: Son
authors.forename: Chang-deok
authors.initials: CD
authors.lastname: Han
authors.forename: Kon Ho
authors.initials: KH
authors.lastname: Lee
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D041883:Chlorobium / Q000201:enzymology*; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D019281:Dimerization; D004926:Escherichia coli / Q000201:enzymology; D008958:Models, Molecular; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D000429:Alcohol Oxidoreductases; C022630:sepiapterin reductase