title: Purification, crystallization and preliminary crystallographic analysis of Arabidopsis thaliana imidazoleglycerol-phosphate dehydratase.
abstract: Imidazoleglycerol-phosphate dehydratase catalyses the sixth step of the histidine-biosynthesis pathway in plants and microorganisms and has been identified as a possible target for the development of novel herbicides. Arabidopsis thaliana IGPD has been cloned and overexpressed in Escherichia coli, purified and subsequently crystallized in the presence of manganese. Under these conditions, the inactive trimeric form of the metal-free enzyme is assembled into a fully active species consisting of a 24-mer exhibiting 432 symmetry. X-ray diffraction data have been collected to 3.0 A resolution from a single crystal at 293 K. The crystal belongs to space group R3, with approximate unit-cell parameters a = b = 157.9, c = 480.0 A, alpha = beta = 90, gamma = 120 degrees and with either 16 or 24 subunits in the asymmetric unit. A full structure determination is under way in order to provide insights into the mode of subunit assembly and to initiate a programme of rational herbicide design.
authors.affiliation: Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, England.
authors.forename: Steven E
authors.initials: SE
authors.lastname: Glynn
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Baker
authors.forename: Svetlana E
authors.initials: SE
authors.lastname: Sedelnikova
authors.forename: Colin W
authors.initials: CW
authors.lastname: Levy
authors.forename: H Fiona
authors.initials: HF
authors.lastname: Rodgers
authors.forename: Jutta
authors.initials: J
authors.lastname: Blank
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Hawkes
authors.forename: David W
authors.initials: DW
authors.lastname: Rice
mesh_terms: D017360:Arabidopsis / Q000201:enzymology*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000378:metabolism*; D003460:Crystallization; D018360:Crystallography, X-Ray; D006836:Hydro-Lyases / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D029681:Arabidopsis Proteins; D006836:Hydro-Lyases; C021434:imidazoleglycerolphosphate dehydratase