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Crystallization and preliminary X‐ray crystallographic analysis of a putative nonribosomal peptide synthase AmbB from Pseudomonas aeruginosa
Author(s) -
Wang Yiwen,
Li Dewang,
Huan Xuelu,
Zhang Lianhui,
Song Haiwei
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14001782
Subject(s) - nonribosomal peptide , escherichia coli , peptide , chemistry , stereochemistry , pseudomonas aeruginosa , monomer , crystallography , molecule , pseudomonas putida , biochemistry , biology , biosynthesis , bacteria , organic chemistry , enzyme , gene , genetics , polymer
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 P 6 1 22 or P 6 5 22, 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.