Biosynthesis of the Proteasome Inhibitor Syringolin A

Ramel, Christina; Tobler, Micha; Meyer, Martin; Bigler, Laurent; Ebert, Marc‐Olivier; Schellenberg, Barbara; Dudler, Robert

doi:10.1201/b13131-15

Cited by 3 publications

(8 citation statements)

References 32 publications

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“…Recent in vivo investigations by Dudler and coworkers into the biosynthetic origin of the ureido-linkage of syringolin A revealed integration of either bicarbonate or carbon dioxide 6. Feeding studies with [ 13 C]-bicarbonate followed by product characterization validated that incorporation was restricted to the carbonyl moiety.…”

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confidence: 99%

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SylC Catalyzes Ureido-Bond Formation During Biosynthesis of the Proteasome Inhibitor Syringolin A

2009

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Syringolins are a class of cyclic tripeptide natural products that are potent and irreversible inhibitors of the eukaryotic proteasome. In addition to being hybrid NRPS/PKS molecules, they also feature an unusual ureido-linkage (red) between two amino acid monomers. Here we report the first in vitro characterization of enzymatic ureido-linkage formation which is catalyzed by an NRPS, SylC. Using 13C- and 18O-labeling studies, we show that biosynthesis occurs via N-carboxylation to form an initial N-carboxy-aminoacyl-S-Ppant enzyme intermediate which undergoes intramolecular cyclization followed by condensation with a second amino acid to form the ureido-containing dipeptide product.

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confidence: 99%

“…The appearance of a [M+1] mass peak by HRMS corroborates the feeding studies of Dudler and coworkers and confirms that SylC alone forms the ureido-linkage from a bicarbonate source (Figure 3B). 6…”

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SylC Catalyzes Ureido-Bond Formation During Biosynthesis of the Proteasome Inhibitor Syringolin A

2009

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“…Syringolin contains an internal ureido motif flanked by two identical amino acids, and the NRPS elongation module SylC is sufficient to catalyse biosynthesis from the amino acid and carbonate through repetitive use of its condensation domain 50. 51 A similar mechanism conceivably applies to pacidamycin biosynthesis.…”

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confidence: 99%

Pacidamycin Biosynthesis: Identification and Heterologous Expression of the First Uridyl Peptide Antibiotic Gene Cluster

et al. 2010

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The pacidamycins are antimicrobial nucleoside antibiotics produced by Streptomyces coeruleorubidus that inhibit translocase I, an essential bacterial enzyme yet to be clinically targeted. The novel pacidamycin scaffold is composed of a pseudopeptide backbone linked by a unique exocyclic enamide to an atypical 3'-deoxyuridine nucleoside. In addition, the peptidyl chain undergoes a double inversion caused by the incorporation of a diamino acid residue and a rare internal ureido moiety. The pacidamycin gene cluster was identified and sequenced, thereby providing the first example of a biosynthetic cluster for a member of the uridyl peptide family of antibiotics. Analysis of the 22 ORFs provided an insight into the biosynthesis of the unique structural features of the pacidamycins. Heterologous expression in Streptomyces lividans resulted in the production of pacidamycin D and the newly identified pacidamycin S, thus confirming the identity of the pacidamycin biosynthetic gene cluster. Identification of this cluster will enable the generation of new uridyl peptide antibiotics through combinatorial biosynthesis. The concise cluster will provide a useful model system through which to gain a fundamental understanding of the way in which nonribosomal peptide synthetases interact.

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“…Our group previously demonstrated that this enzyme is responsible for production of the ureido side-chain and proceeds through a cyclized intermediate. 7 SylD is an NRPS-PKS (polyketide synthase) megasynthase 8 possessing two iterative CAT domains followed by a ketosynthase (KS), dehydratase (DH), acyltransferase (AT), ketoreductase (KR), thiolation (T), and thioesterase (TE) domains. Bioformatic analysis predicts that this protein is responsible for the production and cyclization of the macrolactam core structure.…”

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confidence: 99%

“…SylC is a nonribosomal peptide synthetase (NRPS) composed of a condensation (C), a C-terminal condensation domain (C*), an adenylation (A), and a thiolation (T) domain. Our group previously demonstrated that this enzyme is responsible for production of the ureido side chain and proceeds through a cyclized intermediate . SylD is an NRPS-PKS (polyketide synthase) megasynthase possessing two iterative CAT domains followed by a ketosynthase (KS), dehydratase (DH), acyltransferase (AT), ketoreductase (KR), thiolation (T), and thioesterase (TE) domains.…”

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Enzymatic Timing and Tailoring of Macrolactamization in Syringolin Biosynthesis

et al. 2011

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The enzymatic activation of 3,4-dehydrolysine and subsequent formation of the twelve-membered syringolin macrolactam were investigated. The timing of the desaturation was elucidated through the analysis of the initial adenylation domain of SylD. The SylD-TTE didomain was characterized and demonstrated to be the catalyst for formation of twelve-membered macrocycles. When the SylD thioesterase domain was reacted with a family of acyclic CoA both natural and unnatural macrocycles were generated.

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Biosynthesis of the Proteasome Inhibitor Syringolin A

Cited by 3 publications

References 32 publications

SylC Catalyzes Ureido-Bond Formation During Biosynthesis of the Proteasome Inhibitor Syringolin A

SylC Catalyzes Ureido-Bond Formation During Biosynthesis of the Proteasome Inhibitor Syringolin A

Pacidamycin Biosynthesis: Identification and Heterologous Expression of the First Uridyl Peptide Antibiotic Gene Cluster

Enzymatic Timing and Tailoring of Macrolactamization in Syringolin Biosynthesis

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