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.
The Journal of Antibiotics 08/1973; 26(7):389-90. DOI:10.7164/antibiotics.26.389 · 2.04 Impact Factor
The Journal of Antibiotics 03/1972; 25(2):122-7. DOI:10.7164/antibiotics.25.122 · 2.04 Impact Factor
Article: Solvent content of protein crystals.[Show abstract] [Hide abstract]
ABSTRACT: An analysis has been made, from the data which are currently available, of the solvent content of 116 different crystal forms of globular proteins. The fraction of the crystal volume occupied by solvent is most commonly near 43 %, but has been observed to have values from about 27 to 65%. In many cases this range will be sufficiently restrictive to enable the probable number of molecules in the crystallographic asymmetric unit to be determined directly from the molecular weight of the protein and the space group and unit cell dimensions of the crystal.Journal of Molecular Biology 05/1968; 33(2):491-7. DOI:10.1016/0022-2836(68)90205-2 · 3.96 Impact Factor