Crystallization and preliminary X-ray crystallographic analysis of a putative nonribosomal peptide synthase AmbB from Pseudomonas aeruginosa

• Published in: Acta crystallographica. Section F, Structural biology communications Vol. 70; no. 3; pp. 339 - 342
• Main Authors: Wang, Yiwen, Li, Dewang, Huan, Xuelu, Zhang, Lianhui, Song, Haiwei
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01.03.2014; Wiley Subscription Services, Inc
• Subjects: AmbB; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Chromatography, Affinity; Chromatography, Gel; Crystallization; Crystallization Communications; Crystallography, X-Ray; Peptide Synthases - chemistry; Peptide Synthases - isolation & purification; Pseudomonas aeruginosa; Pseudomonas aeruginosa - enzymology; Pseudomonas aeruginosa; AmbB
• ISSN: 2053-230X; 2053-230X
• DOI: 10.1107/S2053230X14001782

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.