Lasso peptides from proteobacteria: Genome mining employing heterologous expression and mass spectrometry

Hegemann, Julian D.; Zimmermann, Marcel; Zhu, Shaozhou; Klug, Dennis; Marahiel, Mohamed A.

doi:10.1002/bip.22326

Cited by 112 publications

(278 citation statements)

References 54 publications

Supporting

Mentioning

256

Contrasting

Unclassified

Order By: Relevance

“…Firmicute and proteobacterial lasso peptide biosynthetic operons encoding a kinase represent two new types of lasso peptide gene clusters [30,34]. Previously, these cryptic clusters were identified, and the biosynthetic operon of paeninodin was examined to determine how phosphorylated lasso peptides arise in firmicutes [34].…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Dual substrate‐controlled kinase activity leads to polyphosphorylated lasso peptides

et al. 2016

Self Cite

View full text Add to dashboard Cite

Edited by Alfonso ValenciaLasso peptides are characterized by their peculiar lariat knot-like structure. Except for maturation of this fold, post-translational modifications of lasso peptides are rare. However, we recently delineated the biosynthetic pathway of a post-translationally phosphorylated lasso peptide, paeninodin. In this study, further investigation of two kinases revealed their ability to transfer multiple phosphate groups onto precursor peptide substrates, ultimately leading to polyphosphorylated lasso peptides. We found that this polyphosphorylating activity depended on the identity of the phosphate donor and the sequence of the precursor peptide. Our investigations provide new insight into the remarkable strategies for chemical diversification employed by the lasso peptide biosynthetic machinery.

show abstract

Section: Discussionmentioning

confidence: 99%

“…Kinase-encoding operons were observed widely throughout firmicutes and proteobacteria, with gene arrangements clearly divided into two corresponding clades [30,34].…”

Section: In Vitro Polyphosphorylation Of the Native Precursors By Thcmentioning

confidence: 99%

Dual substrate‐controlled kinase activity leads to polyphosphorylated lasso peptides

et al. 2016

Self Cite

View full text Add to dashboard Cite

show abstract

“…This organism has two separate lasso peptide gene clusters, each with an associated isopeptidase gene (15, 16). There have been a large number of lasso peptides discovered in proteobacteria (11,17), and an isopeptidase is commonly associated with such clusters (9).We named the two isopeptidases in A. excentricus AtxE1 and AtxE2. The gene for AtxE1 is found next to the gene cluster that encodes for the biosynthesis of astexin-1, while the AtxE2 gene is located next to the gene cluster encoding astexins-2 and -3 (Fig.…”

mentioning

confidence: 99%

mentioning

confidence: 99%

Elucidating the Specificity Determinants of the AtxE2 Lasso Peptide Isopeptidase

Maksimov

Koos

Zong

et al. 2015

Journal of Biological Chemistry

View full text Add to dashboard Cite

Lasso peptide isopeptidase is an enzyme that specifically hydrolyzes the isopeptide bond of lasso peptides, rendering these peptides linear. To carry out a detailed structure-activity analysis of the lasso peptide isopeptidase AtxE2 from Asticcacaulis excentricus, we solved NMR structures of its substrates astexin-2 and astexin-3. Using in vitro enzyme assays, we show that the C-terminal tail portion of these peptides is dispensable with regards to isopeptidase activity. A collection of astexin-2 and astexin-3 variants with alanine substitutions at each position within the ring and the loop was constructed, and we showed that all of these peptides except for one were cleaved by the isopeptidase. Thus, much like the lasso peptide biosynthetic enzymes, lasso peptide isopeptidase has broad substrate specificity. Quantitative analysis of the cleavage reactions indicated that alanine substitutions in loop positions of these peptides led to reduced cleavage, suggesting that the loop is serving as a recognition element for the isopeptidase.Ribosomally synthesized and post-translationally modified peptides (RiPPs) 3 (1) are a diverse set of natural products that are formed by the action of maturation enzymes on a linear peptide substrate. An emerging theme in the biosynthesis of RiPPs is that the maturation enzymes tend to have broad substrate specificity (2-8). Lasso peptides are a class of RiPPs, characterized by a threaded structure resembling a slipknot (9 -11). The internal macrocycle is realized via a single isopeptide bond installed post-translationally by two maturation enzymes (12). While there is a quickly expanding literature about the lasso peptide maturation enzymes (12-14), there is little known about the catabolism of these molecules. Recently, we reported an enzyme, lasso peptide isopeptidase, that specifically hydrolyzes the isopeptide bond of lasso peptides (Fig. 1A) (15). This enzyme, related to prolyl oligopeptidases, was found in the vicinity of a lasso peptide gene cluster in the freshwater ␣-proteobacterium Asticcacaulis excentricus. This organism has two separate lasso peptide gene clusters, each with an associated isopeptidase gene (15, 16). There have been a large number of lasso peptides discovered in proteobacteria (11,17), and an isopeptidase is commonly associated with such clusters (9).We named the two isopeptidases in A. excentricus AtxE1 and AtxE2. The gene for AtxE1 is found next to the gene cluster that encodes for the biosynthesis of astexin-1, while the AtxE2 gene is located next to the gene cluster encoding astexins-2 and -3 (Fig. 1B) (15). We have previously characterized AtxE2 in vitro (15). This enzyme cleaves astexin-2 and astexin-3, but no crossreactivity was observed between AtxE2 and astexin-1. Whereas astexin-2 and astexin-3 share relatively high sequence homology (identity at 13/24 positions), the astexin-1 sequence is more divergent (Fig. 1C). In addition, we have shown that, at least for astexin-2, the thermally unthreaded variant of the peptide is not a substrate for Atx...

show abstract

“…Aufgrund ihrer Struktur sind Lassopeptide oft gegenüber proteolytischer und chemischer Degradation sowie in manchen Fällen auch thermisch außerge-wçhnlich stabil. [1,3,8,10,14,[16][17][18][19][20][21][22][23] Sie sind aber nicht nur wegen ihrer beispiellosen Faltung und ihrer nichtkovalenten Formstabilisierung von Interesse, sondern auch oft mit interessanten biologischen Eigenschaften verbunden, die von antimikrobiellen [3,10,11,18,20,24,25] über rezeptorantagonistischen [26][27][28] zu inhibitorischen Wirkungen [1,20,24,25,[29][30][31][32] reichen. Da ihre Biosynthesemaschinerie hochpromiskuitiv ist, kçnnen die meisten Reste in ihrer Sequenz mühelos ausgetauscht werden, was ihre erfolgreiche Nutzung zum Aufpropfen von Peptidepitopen mçglich machte.…”

unclassified

Xanthomonine I–III: eine neue Klasse von Lassopeptiden mit einem Makrolactamring aus sieben Aminosäureresten

et al. 2014

Self Cite

View full text Add to dashboard Cite

Lassopeptide gehçren zu den ribosomal synthetisierten und posttranslatorisch modifizierten Peptiden. Ihr gemeinsames Erkennungsmerkmal ist ein N-terminaler Makrolactamring, der von einem C-terminalen Schwanz durchfädelt wird. Diese "Lassofaltung" wird durch sterische Wechselwirkungen aufrechterhalten. In dieser Studie werden die Isolierung und Charakterisierung der Xanthomonine I-III, der ersten Lassopeptide mit einem nur aus sieben Aminosäuren bestehenden Makrolactamring, sowie die Kristallstruktur von Xanthomonin I und die NMR-spektroskopisch ermittelte Struktur von Xanthomonin II beschrieben. Insgesamt wurden 25 Xanthomonin-II-Varianten generiert, um Aspekte der Biosynthese, der Stabilisierung und der Aufrechterhaltug der Lassofaltung zu testen. Diese Mutagenesestudie offenbart die Einschränkungen, die ein so kleiner Ring für das Durchfädeln bedeutet, und zeigt, dass jede Stçpselaminosäure grçßer als Serin eine hitzestabile Lassofaltung in Xanthomonin II aufrechterhalten kann.Lassopeptide sind eine faszinierende Gruppe von Naturstoffen, die das einzigartige Strukturmotiv des Lassoknotens aufweisen. Dieses Strukturmerkmal besteht aus einem Makrolactamring, der zwischen der N-terminalen a-Aminogruppe und der Carbonsäureseitenkette eines Asp-oder GluRests gebildet wird und durch den der C-terminale Schwanz gefädelt ist, welcher durch Aminosäuren mit sterisch anspruchsvollen Seitenketten in Position gehalten wird. [1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16] Diese Aminosäuren, sogenannte Stçpsel, sind über und unter dem Ring positioniert. Aufgrund ihrer Struktur sind Lassopeptide oft gegenüber proteolytischer und chemischer Degradation sowie in manchen Fällen auch thermisch außerge-wçhnlich stabil. [1,3,8,10,14,[16][17][18][19][20][21][22][23] Sie sind aber nicht nur wegen ihrer beispiellosen Faltung und ihrer nichtkovalenten Formstabilisierung von Interesse, sondern auch oft mit interessanten biologischen Eigenschaften verbunden, die von antimikrobiellen [3,10,11,18,20,24,25] über rezeptorantagonistischen [26][27][28] zu inhibitorischen Wirkungen [1,20,24,25,[29][30][31][32] reichen. Da ihre Biosynthesemaschinerie hochpromiskuitiv ist, kçnnen die meisten Reste in ihrer Sequenz mühelos ausgetauscht werden, was ihre erfolgreiche Nutzung zum Aufpropfen von Peptidepitopen mçglich machte.

show abstract

Lasso peptides from proteobacteria: Genome mining employing heterologous expression and mass spectrometry

Cited by 112 publications

References 54 publications

Dual substrate‐controlled kinase activity leads to polyphosphorylated lasso peptides

Dual substrate‐controlled kinase activity leads to polyphosphorylated lasso peptides

Elucidating the Specificity Determinants of the AtxE2 Lasso Peptide Isopeptidase

Xanthomonine I–III: eine neue Klasse von Lassopeptiden mit einem Makrolactamring aus sieben Aminosäureresten

Contact Info

Product

Resources

About