title: Crystallization and preliminary X-ray analysis of molecular chaperone-like diol dehydratase-reactivating factor in ADP-bound and nucleotide-free forms.
abstract: Adenosylcobalamin (coenzyme B12) dependent diol dehydratase (EC 4.2.1.28) catalyzes the conversion of 1,2-diols and glycerol to the corresponding aldehydes. It undergoes mechanism-based inactivation by glycerol. The diol dehydratase-reactivating factor (DDR) reactivates the inactivated holoenzymes in the presence of adenosylcobalamin, ATP and Mg2+ by mediating the release of a damaged cofactor. This molecular chaperone-like factor was overexpressed in Escherichia coli, purified and crystallized in the ADP-bound and nucleotide-free forms by the sandwich-drop vapour-diffusion method. The crystals of the ADP-bound form belong to the orthorhombic system, with space group P2(1)2(1)2(1) and unit-cell parameters a = 83.26, b = 84.60, c = 280.09 A, and diffract to 2.0 A. In the absence of nucleotide, DDR crystals were orthorhombic, with space group P2(1)2(1)2(1) and unit-cell parameters a = 81.92, b = 85.37, c = 296.99 A and diffract to 3.0 A. Crystals of both forms were suitable for structural analysis.
authors.affiliation: Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka, Okayama 700-8530, Japan.
authors.forename: Koichi
authors.initials: K
authors.lastname: Mori
authors.forename: Naoki
authors.initials: N
authors.lastname: Hieda
authors.forename: Mamoru
authors.initials: M
authors.lastname: Yamanishi
authors.forename: Naoki
authors.initials: N
authors.lastname: Shibata
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Toraya
mesh_terms: D000244:Adenosine Diphosphate / Q000737:chemistry*; D000255:Adenosine Triphosphate; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003001:Cloning, Molecular; D003038:Cobamides; D003460:Crystallization / Q000379:methods; D041121:Klebsiella oxytoca / Q000201:enzymology*; D008274:Magnesium; D018832:Molecular Chaperones; D011408:Propanediol Dehydratase / Q000737:chemistry*; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D003038:Cobamides; D018832:Molecular Chaperones; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D011408:Propanediol Dehydratase; C000913:cobamamide; D008274:Magnesium