Open AccessCrystallization and preliminary X‐ray analysis of molecular chaperone‐like diol dehydratase‐reactivating factor in ADP‐bound and nucleotide‐free forms
Author(s) -
Mori Koichi,
Hieda Naoki,
Yamanishi Mamoru,
Shibata Naoki,
Toraya Tetsuo
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105015733
Subject(s) - adenosylcobalamin , orthorhombic crystal system , cofactor , dehydratase , crystallization , chemistry , nucleotide , molecular replacement , crystallography , chaperone (clinical) , glycerol , stereochemistry , guanine , biochemistry , crystal structure , enzyme , organic chemistry , medicine , pathology , gene
Adenosylcobalamin (coenzyme B 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 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 P 2 1 2 1 2 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 P 2 1 2 1 2 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.