scholarly journals DrosophilaEbony: a novel type of nonribosomal peptide synthetase related enzyme with unusually fast peptide bond formation kinetics

FEBS Journal ◽  
2014 ◽  
Vol 281 (22) ◽  
pp. 5147-5158 ◽  
Author(s):  
Silvia Hartwig ◽  
Christine Dovengerds ◽  
Christian Herrmann ◽  
Bernhard T. Hovemann
Keyword(s):  
Bond Formation ◽  
Peptide Bond ◽  
Nonribosomal Peptide Synthetase ◽  
Peptide Bond Formation ◽  
Nonribosomal Peptide ◽  
Formation Kinetics ◽  
Peptide Synthetase ◽  
Related Enzyme

scholarly journals Structural and Functional Insights into a Peptide Bond-Forming Bidomain from a Nonribosomal Peptide Synthetase

Structure ◽  
2007 ◽  
Vol 15 (7) ◽  
pp. 781-792 ◽  
Author(s):  
Stefan A. Samel ◽  
Georg Schoenafinger ◽  
Thomas A. Knappe ◽  
Mohamed A. Marahiel ◽  
Lars-Oliver Essen
Keyword(s):  
Peptide Bond ◽  
Nonribosomal Peptide Synthetase ◽  
Nonribosomal Peptide ◽  
Bond Forming ◽  
Peptide Synthetase

scholarly journals Crystallization and preliminary crystallographic analysis of the first condensation domain of viomycin synthetase

2013 ◽  
Vol 69 (4) ◽  
pp. 412-415 ◽  
Author(s):  
Kristjan Bloudoff ◽  
T. Martin Schmeing
Keyword(s):  
Bond Formation ◽  
Peptide Bond ◽  
Crystallographic Analysis ◽  
Diffusion Method ◽  
Peptide Bond Formation ◽  
Orthorhombic Space Group ◽  
Nonribosomal Peptide ◽  
Cell Parameters ◽  
Peptide Synthetases ◽  
Condensation Domain

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 ofStreptomyces 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 groupP212121, with unit-cell parametersa= 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 Å3 Da−1. Structural determination is in progress.

scholarly journals Peptide Bond Formation in Nonribosomal Peptide Biosynthesis

1998 ◽  
Vol 273 (35) ◽  
pp. 22773-22781 ◽  
Author(s):  
Torsten Stachelhaus ◽  
Henning D. Mootz ◽  
Veit Bergendahl ◽  
Mohamed A. Marahiel
Keyword(s):  
Bond Formation ◽  
Peptide Bond ◽  
Peptide Bond Formation ◽  
Nonribosomal Peptide ◽  
Peptide Biosynthesis

scholarly journals Biosynthesis of Novel Pyoverdines by Domain Substitution in a Nonribosomal Peptide Synthetase of Pseudomonas aeruginosa

2014 ◽  
Vol 80 (18) ◽  
pp. 5723-5731 ◽  
Author(s):  
Mark J. Calcott ◽  
Jeremy G. Owen ◽  
Iain L. Lamont ◽  
David F. Ackerley
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Peptide Bond ◽  
Nonribosomal Peptide Synthetase ◽  
Nonribosomal Peptide ◽  
Content Type ◽  
Threonine Residue ◽  
Peptide Synthetase ◽  
A Domains ◽  
High Yields ◽  
Acid Substrate

ABSTRACTPyoverdine is a fluorescent nonribosomal peptide siderophore made by fluorescent pseudomonads. ThePseudomonas aeruginosanonribosomal peptide synthetase (NRPS) PvdD contains two modules that each incorporate anl-threonine residue at the C-terminal end of pyoverdine. In an attempt to generate modified pyoverdine peptides, we substituted alternative-substrate-specifying adenylation (A) and peptide bond-catalyzing condensation (C) domains into the second module of PvdD. When just the A domain was substituted, the resulting strains produced only wild-type pyoverdine—at high levels if the introduced A domain specified threonine or at trace levels otherwise. The high levels of pyoverdine synthesis observed whenever the introduced A domain specified threonine indicated that these nonnative A domains were able to communicate effectively with the PvdD C domain. Moreover, the unexpected observation that non-threonine-specifying A domains nevertheless incorporated threonine into pyoverdine suggests that the native PvdD C domain exhibited stronger selectivity than these A domains for the incorporated amino acid substrate (i.e., misactivation of a threonine residue by the introduced A domains was more frequent than misincorporation of a nonthreonine residue by the PvdD C domain). In contrast, substitution of both the C and A domains of PvdD generated high yields of rationally modified pyoverdines in two instances, these pyoverdines having either a lysine or a serine residue in place of the terminal threonine. However, C-A domain substitution more commonly yielded a truncated peptide product, likely due to stalling of synthesis on a nonfunctional recombinant NRPS template.

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