Crystallization and preliminary X-ray analysis of molecular chaperone-like diol dehydratase-reactivating factor in ADP-bound and nucleotide-free forms
- Graduate School of Natural Science and Technology, Okayama University, Tsushima-Naka, Okayama 700-8530 (Japan)
- Graduate School of Life Science, University of Hyogo, 3-2-1 Kouto, Ako-gun, Hyogo 678-1297 (Japan)
The molecular chaperone-like reactivating factor for adenosylcobalamin (coenzyme B{sub 12}) dependent diol dehydratase was crystallized in ADP-bound and nucleotide-free forms. Preliminary X-ray analysis indicated that crystals are orthorhombic and diffract to 2.0 Å. Adenosylcobalamin (coenzyme B{sub 12}) 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 Mg{sup 2+} 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{sub 1}2{sub 1}2{sub 1} and unit-cell parameters a = 83.26, b = 84.60, c = 280.09 Å, and diffract to 2.0 Å. In the absence of nucleotide, DDR crystals were orthorhombic, with space group P2{sub 1}2{sub 1}2{sub 1} and unit-cell parameters a = 81.92, b = 85.37, c = 296.99 Å and diffract to 3.0 Å. Crystals of both forms were suitable for structural analysis.
- OSTI ID:
- 22356023
- Journal Information:
- Acta Crystallographica. Section F, Journal Name: Acta Crystallographica. Section F Journal Issue: Pt 6 Vol. 61; ISSN ACSFCL; ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
Similar Records
Crystal structure of substrate free form of glycerol dehydratase
X-ray diffraction analysis of a crystal of HscA from Escherichia coli