Overexpression, crystallization and preliminary X-ray crystallographic analysis of Nudix hydrolase Orf141 from Escherichia coli K-1
- Department of Advanced Technology Fusion, Konkuk University, Hwayang-dong, Gwangjin-gu, Seoul 143-701 (Korea, Republic of)
- Division of Life Science, College of Natural Sciences, Sangmyung University, Hongji-dong 7, Jongno-gu, Seoul 110-743 (Korea, Republic of)
A novel Nudix hydrolase Orf141 from E. coli, which cleaves pyrimidine deoxynucleoside triphosphates, was recently cloned. In order to illuminate the structural and functional features of the novel Nudix hydrolase Orf141, it was expressed, purified and crystallized and a preliminary X-ray crystallographic analysis was conducted. Nudix hydrolases are a family of proteins that contain the characteristic amino-acid sequence GX{sub 5}EX{sub 7}REUXEEXGU (where U is usually I, L or V), the Nudix signature sequence. They catalyze the hydrolysis of a variety of nucleoside diphosphate derivatives such as nucleoside triphosphates, nucleotide sugars, ADP-ribose, dinucleotide coenzymes, diadenosine oligophosphates and capped RNAs. Recently, three new Nudix hydrolases have been found from Escherichia coli; one of them is Orf141, which cleaves pyrimidine deoxynucleoside triphosphates. Orf141 was cloned directly from E. coli K1 strain and was overexpressed in E. coli without any extra residues. Orf141 crystals were successfully obtained using polyethylene glycol 1500 as a precipitant at 285 K. X-ray diffraction data were collected to 3.1 Å resolution using synchrotron radiation. The crystal is a member of the rhombohedral space group H32, with unit-cell parameters a = b = 182.2, c = 62.3 Å, α = 90, β = 90, γ = 120° (hexagonal setting). Two or three monomers are likely to be present in the asymmetric unit, with corresponding V{sub M} values of 2.92 and 1.95 Å{sup 3} Da{sup −1} and solvent contents of 57.9 and 36.9%, respectively.
- OSTI ID:
- 22360552
- Journal Information:
- Acta Crystallographica. Section F, Vol. 63, Issue Pt 9; Other Information: PMCID: PMC2376330; PMID: 17768363; PUBLISHER-ID: pu5197; OAI: oai:pubmedcentral.nih.gov:2376330; Copyright (c) International Union of Crystallography 2007; Country of input: International Atomic Energy Agency (IAEA); ISSN 1744-3091
- Country of Publication:
- United Kingdom
- Language:
- English
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