title: Preparation and crystallization of a Bacillus subtilis arsenate reductase.
abstract: Arsenate reductase (AR) in B. subtilis is encoded by the chromosomal arsC gene. Together with arsB and arsR, arsC participates in detoxification processes for the arsenate and arsenite ions. Full-length arsenate reductase without any modification has been expressed in Escherichia coli and purified in a soluble form. The recombinant protein has been crystallized at 277 K using polyethyleneglycol (PEG) or poly(ethyleneglycol) methyl ether (PME) as the main precipitant. At least two forms of crystals large enough for data collection have been obtained from wild-type protein under different conditions. An orthorhombic crystal diffracted to beyond 2.2 A with space group P2(1)2(1)2(1) and unit-cell parameters a = 51.22, b = 91.62, c = 101.93 A. A near-complete data set has been collected to 2.5 A. The application of the flash-annealing technique was crucial for high resolution during the data collection. The SeMet-substituted AR has also been produced and crystallized under very similar conditions as the wild type, but the unit-cell parameters are very different. The crystals of the SeMet protein diffracted to higher resolution than those of the wild type.
authors.affiliation: Department of Molecular Biophysics, Kemicentrum, PO Box 124, SE-221 00 Lund, Sweden.
authors.forename: Z
authors.initials: Z
authors.lastname: Guan
authors.forename: L
authors.initials: L
authors.lastname: Hederstedt
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.forename: X D
authors.initials: XD
authors.lastname: Su
mesh_terms: D000251:Adenosine Triphosphatases / Q000096:biosynthesis / Q000737:chemistry*; D000595:Amino Acid Sequence; D053501:Arsenite Transporting ATPases; D001412:Bacillus subtilis / Q000201:enzymology*; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D016623:Ion Pumps; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes; D010802:Phylogeny; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
chemical_list: D016623:Ion Pumps; D009097:Multienzyme Complexes; D000251:Adenosine Triphosphatases; D053501:Arsenite Transporting ATPases