Structure and Mechanism of the Lantibiotic Cyclase Involved in Nisin Biosynthesis

Ли, Бо; Yu, John‐Paul J.; Brunzelle, J.S.; Moll, Gert N.; Donk, Wilfred A. van der; Nair, Satish K.

doi:10.1126/science.1121422

Cited by 281 publications

(409 citation statements)

References 31 publications

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“…LanCs have about 400 amino acid residues, and possess a double α-barrel-fold topology (17) and a strictly conserved Cys-Cys-His triad near their C termini for binding of a zinc ion. In vitro reconstitution of the nisin cyclase activity of NisC and solution of its crystal structure have supported a zincdependent mechanism (17,18). The zinc ion is believed to activate the Cys thiols of the precursor peptide for nucleophilic attack on the dehydroamino acids.…”

Section: Resultsmentioning

confidence: 93%

“…This system is a prime example of the highly evolvable nature of ribosomal biosynthesis to access high structural diversity of natural products at low genetic cost. Analysis of the ProcM sequence revealed that this enzyme contains a "CCG" motif (30) rather than a "CHG" motif (17,18) found in all LanCs and most of the LanMs known to date, indicating that ProcM likely uses three Cys residues rather than a Cys-Cys-His triad for binding of the active site zinc ion. Model studies of activation of thiolate nucleophiles by Zn 2+ have demonstrated increased reactivity with an increased number of thiolate ligands (33), suggesting that ProcM may derive its promiscuity in part from a highly active zinc ion (30).…”

Section: Resultsmentioning

confidence: 99%

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Evolution of lanthipeptide synthetases

Zhang

Velásquez

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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173

244

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Lanthionine-containing peptides (lanthipeptides) are a family of ribosomally synthesized and posttranslationally modified peptides containing (methyl)lanthionine residues. Here we present a phylogenomic study of the four currently known classes of lanthipeptide synthetases (LanB and LanC for class I, LanM for class II, LanKC for class III, and LanL for class IV). Although they possess very similar cyclase domains, class II-IV synthetases have evolved independently, and LanB and LanC enzymes appear to not always have coevolved. LanM enzymes from various phyla that have three cysteines ligated to a zinc ion (as opposed to the more common CysCys-His ligand set) cluster together. Most importantly, the phylogenomic data suggest that for some scaffolds, the ring topology of the final lanthipeptides may be determined in part by the sequence of the precursor peptides and not just by the biosynthetic enzymes. This notion was supported by studies with two chimeric peptides, suggesting that the nisin and prochlorosin biosynthetic enzymes can produce the correct ring topologies of epilancin 15X and lacticin 481, respectively. These results highlight the potential of lanthipeptide synthetases for bioengineering and combinatorial biosynthesis. Our study also demonstrates unexplored areas of sequence space that may be fruitful for genome mining. molecular evolution | natural products | phylogeny | posttranslational modification | lantibiotics

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Section: Resultsmentioning

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Evolution of lanthipeptide synthetases

Zhang

Velásquez

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

173

244

View full text Add to dashboard Cite

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“…Recent studies of various RiPP biosynthetic enzymes have provided several examples of biocatalysts that can generate a macrocyclic product from a linear peptide substrate (14,28,30,31,(52)(53)(54). Such enzymes represent a next-generation technological utility, but many of these catalysts are often slow or require sequence constraints on substrates.…”

Section: Discussionmentioning

confidence: 99%

Characterization of the macrocyclase involved in the biosynthesis of RiPP cyclic peptides in plants

Chekan

Estrada

Covello

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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Enzymes that can catalyze the macrocyclization of linear peptide substrates have long been sought for the production of libraries of structurally diverse scaffolds via combinatorial gene assembly as well as to afford rapid in vivo screening methods. Orbitides are plant ribosomally synthesized and posttranslationally modified peptides (RiPPs) of various sizes and topologies, several of which are shown to be biologically active. The diversity in size and sequence of orbitides suggests that the corresponding macrocyclases may be ideal catalysts for production of cyclic peptides. Here we present the biochemical characterization and crystal structures of the plant enzyme PCY1 involved in orbitide macrocyclization. These studies demonstrate how the PCY1 S9A protease fold has been adapted for transamidation, rather than hydrolysis, of acyl-enzyme intermediates to yield cyclic products. Notably, PCY1 uses an unusual strategy in which the cleaved C-terminal follower peptide from the substrate stabilizes the enzyme in a productive conformation to facilitate macrocyclization of the N-terminal fragment. The broad substrate tolerance of PCY1 can be exploited as a biotechnological tool to generate structurally diverse arrays of macrocycles, including those with nonproteinogenic elements.RiPP | biosynthesis | peptide | plant | orbitide

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“…LanC enzymes are zinc metalloproteins in which the bound metal ion activates the thiol moiety of the cysteine residue for nucleophilic addition onto dehydrated S (Dha) and T (Dhb) residues. Consistent with this, PspC contains the conserved H and C residues that coordinate the zinc ion in NisC (44). Thus, PspC is proposed to be the cyclase responsible for (Me)Lan bridge formation in planosporicin.…”

Section: Identification Of P Alba As a Planosporicin Producer Fourmentioning

confidence: 60%

Cloning and Analysis of the Planosporicin Lantibiotic Biosynthetic Gene Cluster of Planomonospora alba

2013

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The increasing prevalence of antibiotic resistance in bacterial pathogens has renewed focus on natural products with antimicrobial properties. Lantibiotics are ribosomally synthesized peptide antibiotics that are posttranslationally modified to introduce (methyl)lanthionine bridges. Actinomycetes are renowned for their ability to produce a large variety of antibiotics, many with clinical applications, but are known to make only a few lantibiotics. One such compound is planosporicin produced by Planomonospora alba, which inhibits cell wall biosynthesis in Gram-positive pathogens. Planosporicin is a type AI lantibiotic structurally similar to those which bind lipid II, the immediate precursor for cell wall biosynthesis. The gene cluster responsible for planosporicin biosynthesis was identified by genome mining and subsequently isolated from a P. alba cosmid library. A minimal cluster of 15 genes sufficient for planosporicin production was defined by heterologous expression in Nonomuraea sp. strain ATCC 39727, while deletion of the gene encoding the precursor peptide from P. alba, which abolished planosporicin production, was also used to confirm the identity of the gene cluster. Deletion of genes encoding likely biosynthetic enzymes identified through bioinformatic analysis revealed that they, too, are essential for planosporicin production in the native host. Reverse transcription-PCR (RT-PCR) analysis indicated that the planosporicin gene cluster is transcribed in three operons. Expression of one of these, pspEF, which encodes an ABC transporter, in Streptomyces coelicolor A3(2) conferred some degree of planosporicin resistance on the heterologous host. The inability to delete these genes from P. alba suggests that they play an essential role in immunity in the natural producer.

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Structure and Mechanism of the Lantibiotic Cyclase Involved in Nisin Biosynthesis

Cited by 281 publications

References 31 publications

Evolution of lanthipeptide synthetases

Evolution of lanthipeptide synthetases

Characterization of the macrocyclase involved in the biosynthesis of RiPP cyclic peptides in plants

Cloning and Analysis of the Planosporicin Lantibiotic Biosynthetic Gene Cluster of Planomonospora alba

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