title: Crystallization and preliminary X-ray crystallographic analysis of a putative nonribosomal peptide synthase AmbB from Pseudomonas aeruginosa.
abstract: AmbB is a putative nonribosomal peptide synthase from Pseudomonas aeruginosa, which is involved in the production of IQS, a potent cell-cell communication signal molecule that integrates the quorum-sensing mechanism and stress response. It consists of 1249 amino acids and contains an AMP-binding domain, a phosphopantetheine-binding (PB) domain and a condensation (C) domain. In this report, a truncated form of AmbB that contains the PB domain and the condensation domain was overexpressed with an N-terminal GST tag in Escherichia coli and purified as a monomer using affinity and size-exclusion chromatography. The recombinant AmbBc (comprising residues 727-1249 of full-length AmbB) was crystallized using the hanging-drop vapour-diffusion method and a full data set was collected to 2.45 Å resolution using a synchrotron-radiation source. The crystals belonged to space group P6122 or P6522, with unit-cell parameters a = b = 87.81, c = 286.8 Å, α = 90, β = 90, γ = 120°, and contained one molecule per asymmetric unit.
authors.affiliation: Life Sciences Institute, Zhejiang University, 388 Yuhangtang Road, Hangzhou, People's Republic of China.
authors.forename: Yiwen
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Life Sciences Institute, Zhejiang University, 388 Yuhangtang Road, Hangzhou, People's Republic of China.
authors.forename: Dewang
authors.initials: D
authors.lastname: Li
authors.affiliation: Institute of Molecular and Cell Biology, 61 Biopolis Drive, Singapore 138673, Singapore.
authors.forename: Xuelu
authors.initials: X
authors.lastname: Huan
authors.affiliation: Institute of Molecular and Cell Biology, 61 Biopolis Drive, Singapore 138673, Singapore.
authors.forename: Lianhui
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Life Sciences Institute, Zhejiang University, 388 Yuhangtang Road, Hangzhou, People's Republic of China.
authors.forename: Haiwei
authors.initials: H
authors.lastname: Song
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000302:isolation & purification; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D010453:Peptide Synthases / Q000737:chemistry* / Q000302:isolation & purification; D011550:Pseudomonas aeruginosa / Q000201:enzymology*
chemical_list: D001426:Bacterial Proteins; D010453:Peptide Synthases; C104385:non-ribosomal peptide synthase
keywords: AmbB; Pseudomonas aeruginosa