title: Crystallization and preliminary crystallographic analysis of the first condensation domain of viomycin synthetase.
abstract: Nonribosomal peptide synthetases (NRPSs) are large multimodular enzymes that synthesize important secondary metabolites such as antibiotics. NRPSs follow a modular synthetic logic whereby each successive amino-acid monomer is added to the peptide chain by successive multi-domain modules. The condensation domain catalyzes the central chemical event in the synthetic cycle, peptide-bond formation, and is present in every elongation module of the NRPS. Viomycin is an antituberculosis nonribosomal peptide that is synthesized by a series of four NRPS proteins and then modified by tailoring proteins. In order to study the mechanisms of peptide-bond formation in viomycin and in NRPSs in general, a structural study of the first condensation domain of the viomycin synthetase protein VioA (VioA-C1) was initiated. The gene for VioA-C1 was cloned from genomic DNA of Streptomyces vinaceus, expressed as an octahistidine-tagged construct and purified by column chromatography. VioA-C1 was crystallized using the sitting-drop vapor-diffusion method. X-ray diffraction data were collected on a rotating-anode source to 2.9 Å resolution. The data could be indexed in the orthorhombic space group P212121, with unit-cell parameters a = 46.165, b = 68.335, c = 146.423 Å. There is likely to be one monomer in the asymmetric unit, giving a solvent content of 49.2% and a Matthews coefficient (VM) of 2.42 Å(3) Da(-1). Structural determination is in progress.
authors.affiliation: Department of Biochemistry, McGill University, Montréal, QC H3G 0B1, Canada.
authors.forename: Kristjan
authors.initials: K
authors.lastname: Bloudoff
authors.forename: T Martin
authors.initials: TM
authors.lastname: Schmeing
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D010453:Peptide Synthases / Q000737:chemistry* / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology*; D014756:Viomycin / Q000096:biosynthesis
chemical_list: D010453:Peptide Synthases; D014756:Viomycin
keywords: NRPS; Streptomyces vinaceus; condensation domain; nonribosomal peptide synthetase; peptide bond formation; peptidyl transferase; viomycin synthetase
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