title: Expression, purification and preliminary crystallographic analysis of the Toxoplasma gondii enoyl reductase.
abstract: The protozoan parasite Toxoplasma gondii is the causative agent of one of the most widespread parasitic infections of man and is a leading cause of congenital neurological birth defects and the third most common cause of food-borne deaths in the United States. Despite this, to date no drugs are available that provide a fully effective treatment. Recently, the antibacterial agent triclosan was shown to inhibit the fatty-acid biosynthesis pathway in T. gondii and to interact with the enoyl reductase (ENR). In order to analyse the potential of triclosan as a lead compound targeting T. gondii ENR and to explore unique features of the apicomplexan enzyme that could be exploited in future drug development, structural studies have been initiated on T. gondii ENR. Crystals of T. gondii ENR in complex with NAD+ and triclosan were grown using the hanging-drop vapour-diffusion method with PEG 8000 as precipitant. The crystals belong to space group P3(2)21, with approximate unit-cell parameters a = 78.1, b = 78.1, c = 188.5 A, alpha = beta = 90, gamma = 120 degrees and a dimer in the asymmetric unit. Test data were collected to beyond 2.6 A on cryocooled crystals (100 K) using a Rigaku MM007 rotating-anode X-ray source, revealing that the crystals are suitable for a full structural determination.
authors.affiliation: The Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, England.
authors.forename: Stephen P
authors.initials: SP
authors.lastname: Muench
authors.forename: Sean T
authors.initials: ST
authors.lastname: Prigge
authors.forename: Liqun
authors.initials: L
authors.lastname: Zhu
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Kirisits
authors.forename: Craig W
authors.initials: CW
authors.lastname: Roberts
authors.forename: Sarah
authors.initials: S
authors.lastname: Wernimont
authors.forename: Rima
authors.initials: R
authors.lastname: McLeod
authors.forename: David W
authors.initials: DW
authors.lastname: Rice
mesh_terms: D000818:Animals; D000981:Antiprotozoal Agents / Q000737:chemistry; D003460:Crystallization / Q000379:methods; D050753:Enoyl-(Acyl-Carrier-Protein) Reductase (NADH) / Q000737:chemistry*; D009243:NAD / Q000737:chemistry; D015800:Protozoan Proteins / Q000737:chemistry; D012997:Solvents; D014122:Toxoplasma / Q000201:enzymology*; D014260:Triclosan / Q000737:chemistry; D014961:X-Ray Diffraction
chemical_list: D000981:Antiprotozoal Agents; D015800:Protozoan Proteins; D012997:Solvents; D009243:NAD; D014260:Triclosan; D050753:Enoyl-(Acyl-Carrier-Protein) Reductase (NADH)