X-domain of peptide synthetases recruits oxygenases crucial for glycopeptide biosynthesis

  title={X-domain of peptide synthetases recruits oxygenases crucial for glycopeptide biosynthesis},
  author={Kristina Haslinger and Madeleine Peschke and Clara Brieke and Eglė Maximowitsch and Max J. Cryle},
Non-ribosomal peptide synthetase (NRPS) mega-enzyme complexes are modular assembly lines that are involved in the biosynthesis of numerous peptide metabolites independently of the ribosome. The multiple interactions between catalytic domains within the NRPS machinery are further complemented by additional interactions with external enzymes, particularly focused on the final peptide maturation process. An important class of NRPS metabolites that require extensive external modification of the… 

Regulation of the P450 Oxygenation Cascade Involved in Glycopeptide Antibiotic Biosynthesis.

A model for P450 recruitment and peptide modification that involves continuous association/dissociation of the P 450 enzymes with the NRPS, followed by specific recognition of the peptide cyclization state by the P450 (scanning) leads to an induced conformational change that enhances the affinity of the enzyme/substrate complex and initiates catalysis.

Structure of the terminal PCP domain of the non‐ribosomal peptide synthetase in teicoplanin biosynthesis

Structurally characterized PCP domains from the final non‐ribosomal peptide synthetase module must play in glycopeptide antibiotic biosynthesis support the hypothesis that it is subtle rearrangements, rather than dramatic conformational changes, which govern carrier protein interactions and selectivity during non-ribosome peptide synthesis.

Structures and function of a tailoring oxidase in complex with a nonribosomal peptide synthetase module

The oxidase is shown to perform dehydrogenation of the thiazoline in the peptide intermediate while it is covalently attached to the NRPS, as the penultimate step in bacillamide D synthesis.

The many faces and important roles of protein-protein interactions during non-ribosomal peptide synthesis.

The current state of understanding of the protein-protein interactions that govern NRPS-mediated biosynthesis is presented, with a focus on insights gained from structural studies relating to CP domain interactions within these impressive peptide assembly lines.

Understanding the early stages of peptide formation during the biosynthesis of teicoplanin and related glycopeptide antibiotics

Re reconstituted modules (M) 1–4 of the NRP synthetase (NRPS) assembly lines that synthesise the clinically relevant type IV GPA teicoplanin and the related compound A40926 indicate that NRPS assembly lines function as dynamic peptide assembly lines and not static megaenzyme complexes, which has significant implications for biosynthesis redesign of these important biosynthetic systems.

The Thioesterase Domain in Glycopeptide Antibiotic Biosynthesis Is Selective for Cross-Linked Aglycones.

GPA TE domains represent another impressive example of the ability of TE domains to act as logic gates during NRPS biosynthesis, ensuring that essential late-stage peptide modifications are completed before catalyzing the release of the mature, bioactive peptide product.

Engineering the indigoidine-synthesising enzyme BpsA for diverse applications in biotechnology

The ability to easily detect synthesis of indigoidine was utilised to provide a versatile reporter to detect a variety of biochemical activities and enable application of BpsA as a reporter for a range of different substrates.

New Structural Data Reveal the Motion of Carrier Proteins in Nonribosomal Peptide Synthesis

It appears as though the A domain alternation alone is insufficient to account for the complete NRPS catalytic cycle and that the loaded state of the PCP must also play a role in choreographing catalysis in these complex and fascinating molecular machines.



Explorations of catalytic domains in non-ribosomal peptide synthetase enzymology.

This review focuses on the recent discoveries and breakthroughs in the structural elucidation, molecular mechanism, and chemical biology underlying the discrete domains within NRPSs.

Phylogenetic analysis of condensation domains in NRPS sheds light on their functional evolution

The reconstruction of the phylogenetic relationship of NRPS C domain subtypes is reported and the sequence motifs of recently discovered subtypes (Dual E/C, DCL and Starter domains) and their characteristic sequence differences are analyzed, mutually and in comparison with LCL domains.

The structure of VibH represents nonribosomal peptide synthetase condensation, cyclization and epimerization domains

The crystal structure of the free-standing NRPS condensation (C) domain VibH, which catalyzes amide bond formation in the synthesis of vibriobactin, a Vibrio cholerae siderophore, is reported.

Cytochrome P450 OxyBtei Catalyzes the First Phenolic Coupling Step in Teicoplanin Biosynthesis

OxyBtei is characterized, the first of the Oxy enzymes in teicoplanin biosynthesis and possesses a structure similar to those of other Oxy proteins and is active in crosslinking NRPS‐bound peptide substrates, but displays a significantly altered activity spectrum compared to its well‐studied vancomycin homologue.

Crystal Structure of the Termination Module of a Nonribosomal Peptide Synthetase

The crystal structure of the 144-kilodalton Bacillus subtilis termination module SrfA-C was solved and has implications for the rational redesign of NRPSs as a means of producing novel bioactive peptides.

Structural Characterization of OxyD, a Cytochrome P450 Involved in β-Hydroxytyrosine Formation in Vancomycin Biosynthesis

A series of residues were identified across known aminoacyl-CP-oxidizing P450s that are highly conserved and cluster in the active site or potential CP binding site of OxyD, allowing sequence based identification of P450 function for this subgroup of P 450s that play vital roles in the biosyntheses of many important natural products in addition to the vancomycin-type antibiotics.

Cytochrome p450sky interacts directly with the nonribosomal peptide synthetase to generate three amino acid precursors in skyllamycin biosynthesis.

P450sky is the first P450 demonstrated to not only interact directly with PCP-bound amino acids within the peptide-forming NRPS but also to do so with three different PCP domains in a specific fashion, representing an expansion of the complexity and scope of NRPS-mediated peptide synthesis.

Structure of the epimerization domain of tyrocidine synthetase A.

The 1.5 Å resolution structure of the cofactor-independent TycA E domain reveals an intimate relationship to the condensation (C) domains of peptide synthetases, implying Glu882 as a candidate acid-base catalyst, whereas His743 stabilizes in the protonated state a transient enolate intermediate of the L↔D isomerization.