title: Crystallization and preliminary X-ray crystallographic studies of NADP-dependent 3-hydroxyisobutyrate dehydrogenase from Thermus thermophilus HB8.
abstract: 3-Hydroxyisobutyrate, a central metabolite in the valine catabolic pathway, is reversibly oxidized to methylmalonate semialdehyde by a specific NADP-dependent dehydrogenase (HIBADH). HIBADH from Thermus thermophilus HB8 has been overexpressed in Escherichia coli and crystallized by the microbatch method using lithium chloride as a precipitant at 296 K. X-ray diffraction data have been collected to 1.80 A resolution at 100 K using synchrotron radiation. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 85.878, b = 106.367, c = 168.639 A. A homotetramer of HIBADH is likely to be present in the asymmetric unit, giving a V(M) of 3.0 A(3) Da(-1) and a solvent content of 59.3%.
authors.affiliation: Highthroughput Factory, RIKEN Harima Institute at SPring-8, 1-1-1 Kouto, Mikazuki-cho, Sayo-gun, Hyogo 679-5148, Japan.
authors.forename: Neratur K
authors.initials: NK
authors.lastname: Lokanath
authors.forename: Ikuya
authors.initials: I
authors.lastname: Shiromizu
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Nodake
authors.forename: Mitsuaki
authors.initials: M
authors.lastname: Sugahara
authors.forename: Shigeyuki
authors.initials: S
authors.lastname: Yokoyama
authors.forename: Seiki
authors.initials: S
authors.lastname: Kuramitsu
authors.forename: Masashi
authors.initials: M
authors.lastname: Miyano
authors.forename: Naoki
authors.initials: N
authors.lastname: Kunishima
mesh_terms: D000429:Alcohol Oxidoreductases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism; D009249:NADP / Q000378:metabolism; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017356:Synchrotrons; D016963:Thermus thermophilus / Q000201:enzymology* / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D009249:NADP; D000429:Alcohol Oxidoreductases; C021398:3-hydroxyisobutyrate dehydrogenase