Structure and Biosynthesis of the Jamaicamides, New Mixed Polyketide-Peptide Neurotoxins from the Marine Cyanobacterium Lyngbya majuscula

Edwards, Daniel; Márquez, Brian L.; Nogle, Lisa; McPhail, Kerry L.; Goeger, Douglas E.; Roberts, M. W.; Gerwick, William H.

doi:10.1016/j.chembiol.2004.03.030

Cited by 467 publications

(565 citation statements)

References 67 publications

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“…This means that the bromination reaction either takes place while the substrate is attached to JamC or otherwise on one of the other NRPS or PKS proteins found on the jamaicamide biosynthetic pathway or after the jamaicamide is fully assembled. 5-Hexanoic acid and 5-hexynoic acid were also alternative substrates, in agreement with the classical pyrophosphate exchange assay (39).…”

Section: Discussionsupporting

confidence: 84%

“…To illustrate that this method was not only valid for NikP1, the substrate loading for the following systems were examined: the CloN5/CloN4 carrier/adenylation domain pair involved in the formation of clorobiocin (Table 1) (5), CouN5/CouN4 involved in the formation of the antibiotic coumermycin (5), EntB(ArCP)/EntE involved in the formation of the siderophore enterobactin (24), HMWP2, involved in the formation of the siderophore yersiniabactin (27), PchE/PchD involved in the biosynthesis of the siderophore pyochelin (38), JamC/JamA involved in the formation of the neurotoxin jamaicamide (39), and two NRPS modules from the orphan NRPS/PKS gene cluster on Bacillus subtilis for which the product is unknown (40,41,42). The adenylation-carrier di-domains analyzed are from PksN (BG12652) and the second NRPS module from PksJ (BG10929, gene annotation from http://genolist.pasteur.fr/SubtiList/.…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Activity Screening of Carrier Domains within Nonribosomal Peptide Synthetases Using Complex Substrate Mixtures and Large Molecule Mass Spectrometry

et al. 2006

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For screening a pool of potential substrates that load carrier domains found in non-ribosomal peptide synthetases, large molecule mass spectrometry is shown to be an ideal, unbiased assay. Combining the high resolving power of Fourier-Transform Mass Spectrometry with the ability of adenylation domains to select their own substrates, the mass change that takes place upon formation of a covalent intermediate thus identifies the substrate. This assay has an advantage over traditional radiochemical assays in that many substrates, the substrate pool, can be screened simultaneously. Using proteins on the nikkomycin, clorobiocin, coumermycin A 1 , yersiniabactin, pyochelin and enterobactin biosynthetic pathways as proof-of-principle, preferred substrates are readily identified from substrate pools. Furthermore this assay can be used to provide insight into the timing of tailoring events of biosynthetic pathways as demonstrated using the bromination reaction found on the jamaicamide biosynthetic pathway. Finally, this assay can provide insight into the role and function of orphan gene clusters for which the encoded natural product is unknown. This is demonstrated by identifying the substrates for two NRPS modules from the genes pksN and pksJ, that are found on an orphan NRPS/PKS hybrid cluster from Bacillus subtilis. This new assay format is especially timely for activity screening in an era when new types of thiotemplate assembly lines that defy classification are being discovered at an accelerating rate. KeywordsElectrospray Fourier transform mass spectrometry; Non-ribosomal peptide synthetase; Polyketide synthase; Carrier domains; Post-translational modification Almost weekly, new non-ribosomal peptide synthetase (NRPS) gene clusters encoding for proteins that biosynthesize bioactive compounds are discovered (1-4). Since many of the compounds produced by non-ribosomal peptide synthetase (NRPS) as well as polyketide * To whom correspondence should be addressed, kelleher@scs.uiuc.edu, fax number 217 244-8068 and phone number 217 NIH Public Access Author ManuscriptBiochemistry. Author manuscript; available in PMC 2008 October 9. NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript synthase (PKS) paradigm have potent medicinal utility or are involved in virulence, and display unusual chemistry, they are of great academic and industrial interest. Therefore we wanted to develop an alternative method for substrate screening to complement the more traditional radioactive assays (5,6). In NRPS and PKS systems, the substrates and intermediates are loaded onto and processed while attached to the pantetheinyl functionality on a carrier domain (7,8).Examples of NRPS or hybrid NRPS/PKS natural products for which substrates load onto carrier domains are: the siderophore pyoverdine (9), a virulence factor excreted by pseudomonads, the antimicrobial agents penicillin (10), vancomycin (11) gramicidin (12) and the antitumor agent calicheamycin (13). With ∼300 genomes sequenced and available in the public ...

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Section: Discussionsupporting

confidence: 84%

Section: Resultsmentioning

confidence: 99%

Activity Screening of Carrier Domains within Nonribosomal Peptide Synthetases Using Complex Substrate Mixtures and Large Molecule Mass Spectrometry

et al. 2006

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“…In addition to the HMGS-type gene pedP, several other genes were identified that putatively participate in exomethylene bond biosynthesis. Evidence for the role of PedL, PedM, and PedN in this pathway comes from the comparison of gene clusters involved in the biosynthesis of polyketides with ␤-branches, including mupirocin (23), the antibiotic TA (33), leinamycin (34), and the jamaicamides (35). All gene clusters contain a conserved, clustered set of genes encoding an HMGS, a variable number of CR genes (usually described as enoyl-CoA hydratases but most likely catalyzing different reactions), and a monofunctional KS and ACP.…”

Section: Discussionmentioning

confidence: 99%

Antitumor polyketide biosynthesis by an uncultivated bacterial symbiont of the marine sponge Theonella swinhoei

Piel

Hui

Wen

et al. 2004

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

502

405

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Bacterial symbionts have long been suspected to be the true producers of many drug candidates isolated from marine invertebrates. Sponges, the most important marine source of biologically active natural products, have been frequently hypothesized to contain compounds of bacterial origin. This symbiont hypothesis, however, remained unproven because of a general inability to cultivate the suspected producers. However, we have recently identified an uncultured Pseudomonas sp. symbiont as the most likely producer of the defensive antitumor polyketide pederin in Paederus fuscipes beetles by cloning the putative biosynthesis genes. Here we report closely related genes isolated from the highly complex metagenome of the marine sponge Theonella swinhoei, which is the source of the onnamides and theopederins, a group of polyketides that structurally resemble pederin. Sequence features of the isolated genes clearly indicate that it belongs to a prokaryotic genome and should be responsible for the biosynthesis of almost the entire portion of the polyketide structure that is correlated with antitumor activity. Besides providing further proof for the role of the related beetle symbiont-derived genes, these findings raise intriguing ecological and evolutionary questions and have important general implications for the sustainable production of otherwise inaccessible marine drugs by using biotechnological strategies. F or almost 30 years, symbiotic bacteria have been discussed as the likely true producers of numerous natural products isolated from marine invertebrates (1). Particularly suggestive of a bacterial origin is the similarity of many compounds from sponges, tunicates, or bryozoans to complex polyketides and nonribosomal peptides, two groups of metabolites that are otherwise known exclusively from microorganisms (2, 3). Into these structural families fall some of today's most promising drug candidates, such as discodermolide, bryostatin 1, aplidine, and ET-743 (4). The existence of producing symbionts could have far reaching consequences for the development of sustainable, fermentation-based sources of invertebrate-derived drug candidates, almost all of which are currently inaccessible in large amounts. Several studies have tried to pinpoint the producers by using cultivation (5), cell separation (6), immunolocalization (7), or in situ hybridization (8) approaches. However, as none of these methods have so far provided unambiguous results, the true origin of the compounds still remains an enigma. We have recently used a genetic strategy to provide insights into natural product symbiosis in terrestrial insects (9). Rove beetles of the genera Paederus and Paederidus are the source of the highly active antitumor polyketide pederin (Fig. 1), which they use as chemical defense (10). We isolated a set of putative pederin biosynthesis genes (9, 11) from the metagenome of Paederus fuscipes beetles and traced them to a bacterial symbiont with the closest relationship to Pseudomonas aeruginosa (12). The genes encode a mixed modul...

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“…An analysis of conserved catalytic motifs in the related methyltransferases shows that none of the metazoan FASs contains an intact set of catalytic residues, in agreement with the observation that these enzymes do not methylate their products and are not SAM-dependent. However, in related fungal and bacterial polyketide synthases, intact homologues of this domain are found in equivalent positions and are leading to the production of (partially) methylated products (Edwards et al 2004 ;Fujii et al 2005 ;Molnar et al 2000). The significance of the yME domain for animal FAS remains unclear ; it is probably a remnant from an evolutionary precursor multienzyme producing branched fatty acids or polyketides, and it may still play a steric or stabilizing role for the overall functioning of the enzyme.…”

Section: Domain Structuresmentioning

confidence: 99%

Structure and function of eukaryotic fatty acid synthases

Maier

Leibundgut

Boehringer

et al. 2010

Quart. Rev. Biophys.

119

127

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Abstract. In all organisms, fatty acid synthesis is achieved in variations of a common cyclic reaction pathway by stepwise, iterative elongation of precursors with two-carbon extender units. In bacteria, all individual reaction steps are carried out by monofunctional dissociated enzymes, whereas in eukaryotes the fatty acid synthases (FASs) have evolved into large multifunctional enzymes that integrate the whole process of fatty acid synthesis. During the last few years, important advances in understanding the structural and functional organization of eukaryotic FASs have been made through a combination of biochemical, electron microscopic and X-ray crystallographic approaches. They have revealed the strikingly different architectures of the two distinct types of eukaryotic FASs, the fungal and the animal enzyme system. Fungal FAS is a 2Á6 MDa a 6 b 6 heterododecamer with a barrel shape enclosing two large chambers, each containing three sets of active sites separated by a central wheel-like structure. It represents a highly specialized micro-compartment strictly optimized for the production of saturated fatty acids. In contrast, the animal FAS is a 540 kDa X-shaped homodimer with two lateral reaction clefts characterized by a modular domain architecture and large extent of conformational flexibility that appears to contribute to catalytic efficiency.

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Structure and Biosynthesis of the Jamaicamides, New Mixed Polyketide-Peptide Neurotoxins from the Marine Cyanobacterium Lyngbya majuscula

Cited by 467 publications

References 67 publications

Activity Screening of Carrier Domains within Nonribosomal Peptide Synthetases Using Complex Substrate Mixtures and Large Molecule Mass Spectrometry

Activity Screening of Carrier Domains within Nonribosomal Peptide Synthetases Using Complex Substrate Mixtures and Large Molecule Mass Spectrometry

Antitumor polyketide biosynthesis by an uncultivated bacterial symbiont of the marine sponge Theonella swinhoei

Structure and function of eukaryotic fatty acid synthases

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