Iterative acylation on mature lasso peptides by widespread acetyltransferases for lipolasso production

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2025, Номер unknown

Опубликована: Янв. 1, 2025

Abstract The biosynthesis of ribosomally synthesized and post-translationally modified peptides (RiPPs) leverages iterative catalysis to enhance structural biological diversity. Traditionally, enzymes install multiple post-translational modifications (PTMs) on linear peptides, rather than mature RiPPs with intricate three-dimensional structures, which would require complex changes in substrate binding poses. Here, we present a prolific class GCN5-related N-acetyltransferases (GNATs) that iteratively consecutively acylate two Lys residues within the loop ring motifs lasso diverging from typical modification peptides—an unprecedented function for PTM enzymes. Utilizing high-resolution cryo-electron microscopy enzymatic reconstitution, mapped peptide pocket IatT pinpointed key involved demarcating distinct acetylation steps. Structure-based engineering IatT’s acetyl group recognition site expanded cavity accommodate longer-chain acyl groups, leading creation lipolasso novel ribosomal lipopeptide. This strategy can be applied any RiPP BGC encoding GNAT, facilitating efficient diversification rare lipopeptides. Graphic

Язык: Английский

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An integrated platform for decoding hydrophilic peptide fingerprints of hepatocellular carcinoma using artificial intelligence and two-dimensional nanosheets

Journal of Materials Chemistry B, Год журнала: 2024, Номер 12(31), С. 7532 - 7542

Опубликована: Янв. 1, 2024

Hydrophilic peptides (HPs) play a critical role in the pathogenesis of hepatocellular carcinoma (HCC). However, comprehensive and in-depth high-throughput analysis specific changes HPs associated with HCC remains unrealized, due to complex nature biological fluids challenges mining patterns large data sets. The clinical diagnosis still lacks non-destructive accurate classification method, given limited specificity widely used biomarkers. To address these challenges, we have established multifunctional platform that integrates artificial intelligence computation, hydrophilic interaction extraction HPs, MALDI-MS testing. This aims achieve highly sensitive HP fingerprinting for HCC. method not only facilitates efficient detection but also achieves remarkable 100.00% diagnostic accuracy test cohort, supported by machine learning algorithms. By constructing panel 10 characteristic features, achieved 98% cohort rapid identified 62 deeply involved pathways related liver diseases. integrated strategy provides new research directions future biomarker studies as well early individualized treatment

Язык: Английский

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Characterization of an Iterative Halogenase Acting on Ribosomal Peptides Underlies the Combinatorial Biosynthesis Logic of Lasso Peptides

Journal of Natural Products, Год журнала: 2025, Номер unknown

Опубликована: Фев. 20, 2025

Halogenation is commonly utilized in medicinal chemistry for the improvement of drug leads. Flavin-dependent halogenases (FDHs) are ubiquitous across all domains life, yet iterative FDHs rare biosynthesis ribosomally synthesized and post-translationally modified peptides (RiPPs). Herein, we characterize a novel FDH, ChlH, which orchestrates nonsequential chlorination two specific Trp within core peptide lasso precursor containing three Trp. Biochemical computational studies enable characterization employs unique protein-peptide interactions (PPIs) between its distinct N- C-terminal motifs crucial recognition sequence (RS-II) downstream RS-I leader peptide. Previous have demonstrated indispensability biosynthesis, while RS-II was considered to be replaceable. Furthermore, find that substantially contributes PPI. Bioinformatic analysis reveals prevalence homologous biosynthetic gene clusters (BGCs) various RiPP classes. Heterologous expression chl BGC yields non-, mono-, dichlorinated peptides, with chlorination, particularly dichlorination, enhancing their antibacterial activity. This study expands FDH activity spectrum include catalysis on ribosomal underscores significance tailoring enzymes combinatorial peptides.

Язык: Английский

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A Bacterial Sulfotransferase Catalyzes an Unusual Di‐Sulfation in Natural Products Biosynthesis

ChemBioChem, Год журнала: 2025, Номер unknown

Опубликована: Апрель 4, 2025

Sulfation is a widely used strategy in nature to modify the solubility, polarity, and biological activities of molecules. The enzymes catalyzing sulfation, sulfotransferases (STs), are typically highly specific single sulfation site molecule. Herein, identification characterization sulfated adipostatins reported reveals novel sulfotransferase, AdpST, which responsible for di-sulfation at two sites adipostatins. initial bioinformatic analysis search adipostatin analogs from Streptomyces davaonensis DSM101723 identifies adpST 3'-phosphoadenosine-5'-phosphosulfate (PAPS) biosynthetic cassette, co-clustered with adipostatin-encoding type III polyketide synthase. Mono- di-sulfated discovered extracts S. DSM101723, whereas bacterial natural products has not been reported. Using series vivo vitro experiments, it confirmed that AdpST solely both mono- adipostatins, catalytic activity identified PAPS-dependent STs date. It further demonstrated dedicated PAPS cassette improves capacity. Lastly, determined shares similarity small group uncharacterized STs, suggesting presence additional unique nature, phylogenetically distant many characterized STs.

Язык: Английский

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Deciphering the chemical landscape and potential ecological function of RiPPs from the untapped Archaea domain

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Окт. 8, 2024

Abstract Chemical communication is crucial in ecosystems with complex microbial communities. However, the difficulties inherent to cultivation of archaea have led a limited understanding their chemical language, especially regarding structure diversity and function secondary or specialized metabolites (SMs). Our comprehensive investigation into biosynthetic potential archaea, combined metabolic analyses first report heterologous expression has unveiled previously unexplored capabilities archaeal ribosomally synthesized post-translationally modified peptides (RiPPs). We identified twenty-four new lanthipeptides RiPPs exhibiting unique characteristics, including novel subfamily featuring an type diamino-dicarboxylic (DADC) termini, largely expanding landscape SMs. This sheds light on novelty emphasizes as untapped resource for natural product discovery. Additionally, demonstrate specific antagonistic activity against haloarchaea, mediating biotic interaction halophilic niche. Furthermore, they showcase ecological role enhancing host’s motility by inducing rod-shaped cell morphology upregulating archaellin gene expression, facilitating abiotic environments. These discoveries broaden our language provide promising prospects future exploration SM-mediated interaction. Figure

Язык: Английский

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