Insights into the Biosynthesis and Stability of the Lasso Peptide Capistruin

Knappe, Thomas A.; Linne, Uwe; Robbel, Lars; Marahiel, Mohamed A.

doi:10.1016/j.chembiol.2009.11.009

Cited by 113 publications

(184 citation statements)

References 24 publications

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“…There is increasing evidence that lasso peptides are highly redesignable (39). Multiple amino acid substitutions can be tolerated in MccJ25, particularly in its loop (40,41), and single and some double substitutions are tolerated within the ring and tail portion of capistruin (42). The large size and high polarity of astexin-1 make it attractive as a scaffold in which both the loop and tail regions can be modified to generate lasso peptides with new functions.…”

Section: Discussionmentioning

confidence: 99%

Precursor-centric genome-mining approach for lasso peptide discovery

Maksimov¹,

Pelczer²,

Link

2012

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

135

232

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Lasso peptides are a class of ribosomally synthesized posttranslationally modified natural products found in bacteria. Currently known lasso peptides have a diverse set of pharmacologically relevant activities, including inhibition of bacterial growth, receptor antagonism, and enzyme inhibition. The biosynthesis of lasso peptides is specified by a cluster of three genes encoding a precursor protein and two enzymes. Here we develop a unique genome-mining algorithm to identify lasso peptide gene clusters in prokaryotes. Our approach involves pattern matching to a small number of conserved amino acids in precursor proteins, and thus allows for a more global survey of lasso peptide gene clusters than does homology-based genome mining. Of more than 3,000 currently sequenced prokaryotic genomes, we found 76 organisms that are putative lasso peptide producers. These organisms span nine bacterial phyla and an archaeal phylum. To provide validation of the genome-mining method, we focused on a single lasso peptide predicted to be produced by the freshwater bacterium Asticcacaulis excentricus. Heterologous expression of an engineered, minimal gene cluster in Escherichia coli led to the production of a unique lasso peptide, astexin-1. At 23 aa, astexin-1 is the largest lasso peptide isolated to date. It is also highly polar, in contrast to many lasso peptides that are primarily hydrophobic. Astexin-1 has modest antimicrobial activity against its phylogenetic relative Caulobacter crescentus. The solution structure of astexin-1 was determined revealing a unique topology that is stabilized by hydrogen bonding between segments of the peptide.bioinformatics | protein NMR

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Section: Discussionmentioning

confidence: 99%

Precursor-centric genome-mining approach for lasso peptide discovery

Maksimov¹,

Pelczer²,

Link

2012

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

135

232

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“…In both peptides, the steric entrapping of the C-terminal tail in the macrolactam ring relies on the presence of a bulky amino acid located below the ring: Y20 for MccJ25 [20] and R15 for capistruin [21]. In addition, both peptides contain a bulky amino acid located above the ring: F19 for MccJ25 and R11 for capistruin.…”

Section: G N W H G T a P D W F F N Y Y W G F I G W G N D I F G H Y S mentioning

confidence: 99%

Topoisomer Differentiation of Molecular Knots by FTICR MS: Lessons from Class II Lasso Peptides

Zirah

Afonso

Linne

et al. 2011

J. Am. Soc. Mass Spectrom.

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Lasso peptides constitute a class of bioactive peptides sharing a knotted structure where the C-terminal tail of the peptide is threaded through and trapped within an N-terminal macrolactam ring. The structural characterization of lasso structures and differentiation from their unthreaded topoisomers is not trivial and generally requires the use of complementary biochemical and spectroscopic methods. Here we investigated two antimicrobial peptides belonging to the class II lasso peptide family and their corresponding unthreaded topoisomers: microcin J25 (MccJ25), which is known to yield two-peptide product ions specific of the lasso structure under collisioninduced dissociation (CID), and capistruin, for which CID does not permit to unambiguously assign the lasso structure. The two pairs of topoisomers were analyzed by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS) upon CID, infrared multiple photon dissociation (IRMPD), and electron capture dissociation (ECD). CID and ECD spectra clearly permitted to differentiate MccJ25 from its non-lasso topoisomer MccJ25-Icm, while for capistruin, only ECD was informative and showed different extent of hydrogen migration (formation of c•/z from c/z • ) for the threaded and unthreaded topoisomers. The ECD spectra of the triply-chargedMccJ25 and MccJ25-lcm showed a series of radical b-type product ions ðb 0 I n Þ. We proposed that these ions are specific of cyclic-branched peptides and result from a dual c/z • and y/b dissociation, in the ring and in the tail, respectively. This work shows the potentiality of ECD for structural characterization of peptide topoisomers, as well as the effect of conformation on hydrogen migration subsequent to electron capture.

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“…Similarly, the hydrogen atoms of the molecular axle located on the other side of the molecular barrier H 11 and H 12 are both shielded by the BMP25C8: actually, this latter lies on the triazolium moiety and its aromatic rings can reach the other side of the molecular barrier (respectively Δδ = −0.23, −0.41 ppm). On the contrary, the hydrogen atoms located between the aniline and the triazolium unit H 16 , H 17 , H 18 and H 19 and in a much lesser extent H 13 Figures 6 and 7). The ROESY 1 H-NMR experiment carried out on the protonated lasso ( Figure 6) provided the evidences of the lasso architecture with a localization of the BMP25C8 around the anilinium station (relevant correlation peaks highlighted in cyan).…”

Section: Studies Of the Equilibrium Between The Unthreaded Compound 1mentioning

confidence: 99%

“…In view of their structural-dependent properties, lasso peptides could be considered as potential scaffold for therapeutic peptides [17]. With this aim, several biosyntheses and structure-activity analysis of a wide range of lasso peptides were realized [18][19][20]. Recently, Marahiel et al have highlighted the interest of incorporating a RGD peptide sequence in the turn of the lasso MccJ25 [21].…”

Section: Introductionmentioning

confidence: 99%

A pH-Sensitive Peptide-Containing Lasso Molecular Switch

2013

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Abstract:The synthesis of a peptide-containing lasso molecular switch by a selfentanglement strategy is described. The interlocked [1] rotaxane molecular machine consists of a benzometaphenylene [25]crown-8 (BMP25C8) macrocycle surrounding a molecular axle. This molecular axle contains a tripeptidic sequence and two molecular stations: a N-benzyltriazolium and a pH-sensitive anilinium station. The tripeptide is located between the macrocycle and the triazolium station, so that its conformation can be tailored depending on the shuttling of the macrocycle from one station to the other. At acidic pH, the macrocycle resides around the anilinium moiety, whereas it shuttles around the triazolium station after deprotonation. This molecular machinery thus forces the lasso to adopt a tightened or a loosened conformation.

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Insights into the Biosynthesis and Stability of the Lasso Peptide Capistruin

Cited by 113 publications

References 24 publications

Precursor-centric genome-mining approach for lasso peptide discovery

Precursor-centric genome-mining approach for lasso peptide discovery

Topoisomer Differentiation of Molecular Knots by FTICR MS: Lessons from Class II Lasso Peptides

A pH-Sensitive Peptide-Containing Lasso Molecular Switch

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