ACS Chemical Biology最新文献

{"title":"High-Throughput Activity Reprogramming of Proteases (HARP)","authors":"Samantha G. Martinusen,&nbsp;, ,&nbsp;Ethan W. Slaton,&nbsp;, ,&nbsp;Seyednima Ajayebi,&nbsp;, ,&nbsp;Marian A. Pulgar,&nbsp;, ,&nbsp;Cassidy F. Simas,&nbsp;, ,&nbsp;Sage E. Nelson,&nbsp;, ,&nbsp;Amit Dutta,&nbsp;, ,&nbsp;Julia T. Besu,&nbsp;, ,&nbsp;Steven Bruner,&nbsp;, and ,&nbsp;Carl A. Denard*,&nbsp;","doi":"10.1021/acschembio.5c00230","DOIUrl":"10.1021/acschembio.5c00230","url":null,"abstract":"<p >Developing potent and selective protease inhibitors remains a grueling, iterative, and often unsuccessful endeavor. Although macromolecular inhibitors can achieve single-enzyme specificity, platforms used for macromolecular inhibitor discovery are optimized for high-affinity binders, requiring extensive downstream biochemical characterization to isolate rare inhibitors. Here, we developed the High-throughput Activity Reprogramming of Proteases (HARP) platform. HARP is a yeast-based functional screen that isolates protease-inhibitory macromolecules from large libraries by coupling their inhibition of endoplasmic reticulum-resident proteases to a selectable phenotype on the cell surface. Endowed with high dynamic range and resolution, HARP enabled the isolation of low-nanomolar-range inhibitory nanobodies against tobacco etch virus protease and human kallikrein 6, including a rare 10.5 nM <i>K</i>_<i>I</i> TEVp uncompetitive inhibitor. Structural modeling and deep sequencing all provide insights into the molecular determinants of inhibitors and reinforce HARP’s foundational findings. Overall, HARP is a premier platform for discovering modulatory macromolecules from various synthetic scaffolds against enzyme targets.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2381–2392"},"PeriodicalIF":3.8,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145123771","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Phylogenomic Identification of a Highly Conserved Copper-Binding RiPP Biosynthetic Gene Cluster in Marine Microbulbifer Bacteria","authors":"Yifan Tang,&nbsp;, ,&nbsp;Weimao Zhong,&nbsp;, ,&nbsp;Longping Fu,&nbsp;, ,&nbsp;Emmanuel Asante,&nbsp;, ,&nbsp;Anastasiia Kostenko,&nbsp;, ,&nbsp;F. N. U. Vidya,&nbsp;, ,&nbsp;Paige Mandelare-Ruiz,&nbsp;, ,&nbsp;Tamilore T. Adeogun,&nbsp;, ,&nbsp;Gabriel P. Anderson,&nbsp;, ,&nbsp;Benjamin E. Edmonds,&nbsp;, ,&nbsp;Oscar Fang,&nbsp;, ,&nbsp;Michelle Han,&nbsp;, ,&nbsp;Alia S. Hollingsworth,&nbsp;, ,&nbsp;Amna R. Ingham,&nbsp;, ,&nbsp;Carlyn R. Kirby,&nbsp;, ,&nbsp;Alice Landrum,&nbsp;, ,&nbsp;Connor R. Mack,&nbsp;, ,&nbsp;Nikki S. Nobari,&nbsp;, ,&nbsp;Emma J. Oswald,&nbsp;, ,&nbsp;Cecilia L. Polevoy,&nbsp;, ,&nbsp;Yasmin Sharifian,&nbsp;, ,&nbsp;Timothy J. So,&nbsp;, ,&nbsp;Joelee R. Stokes,&nbsp;, ,&nbsp;Reniya S. Thompson,&nbsp;, ,&nbsp;Rishabh Vuthamaraju,&nbsp;, ,&nbsp;Elaine C. Wang,&nbsp;, ,&nbsp;William H. Yang,&nbsp;, ,&nbsp;Alison E. Onstine,&nbsp;, ,&nbsp;Valerie J. Paul,&nbsp;, ,&nbsp;Ronghu Wu,&nbsp;, ,&nbsp;Allegra T. Aron,&nbsp;, and ,&nbsp;Vinayak Agarwal*,&nbsp;","doi":"10.1021/acschembio.5c00507","DOIUrl":"10.1021/acschembio.5c00507","url":null,"abstract":"<p >Conserved biosynthetic gene clusters (BGCs) are often tied to the production of natural products that perform critical functions in an organism’s physiology and ecological interactions. Here, by phylogenetic analysis across the bacterial genus, we report the obligate conservation of a BGC in genomes of cosmopolitan marine <i>Microbulbifer</i> bacteria. This genus is a common member of marine microbiomes, and this BGC was conserved in <i>Microbulbifer</i> genomes regardless of phylogenetic or geographical dispersal. The post-translationally modified peptidic product encoded by this BGC─which was accessed via heterologous production and its structure elucidated using a combination of mass spectrometry and NMR spectroscopy─was found to be a copper chelator. Similar BGCs were then found in genomes of other marine bacterial genera coinhabiting the microbiomes of sponges and corals. The phylogenomic workflows described herein were implemented in a pedagogic setting at the Georgia Institute of Technology to provide hands-on instruction to undergraduate students in bacterial phylogeny, genome mining, and natural product chemistry.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2462–2474"},"PeriodicalIF":3.8,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acschembio.5c00507","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145084514","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Chemical Probes that Target a Dissociative LuxR-Type Quorum Sensing Receptor in Gram-Negative Bacteria","authors":"Irene M. Stoutland,&nbsp;, ,&nbsp;Guadalupe Aguirre-Figueroa,&nbsp;, and ,&nbsp;Helen E. Blackwell*,&nbsp;","doi":"10.1021/acschembio.5c00490","DOIUrl":"10.1021/acschembio.5c00490","url":null,"abstract":"<p >Quorum sensing (QS) allows bacteria to respond to changes in cell density and participate in collective behaviors. Interfering with QS could provide a strategy to block pathogenicity, reduce biofouling, and support biotechnology. Many common Gram-negative bacteria use LuxR-type QS receptors that regulate gene transcription in response to <i>N-</i>acyl <span>l</span>-homoserine lactone (AHL) signals. The most-studied LuxR-type receptors operate via an associative mechanism, i.e., they dimerize and associate with DNA upon ligand binding. In contrast, members of the less-studied class of dissociative LuxR-type receptors bind DNA as dimers in the absence of a ligand and dissociate from DNA upon ligand binding. Few chemical tools to modulate dissociative receptor activity are known. Such probes could provide new entry into mechanistic studies of LuxI/LuxR-type QS in general. In this report, we describe the discovery of synthetic modulators of EsaR, a dissociative LuxR-type receptor present in the plant pathogen <i>Pantoea stewartii</i>, based on AHL scaffolds. Compound activity was evaluated using both cell-based EsaR reporters and a phenotypic assay. We identified compound features associated with agonistic activity in EsaR, some of which were comparable to those of synthetic ligands active in other LuxR-type receptors. However, in contrast to prior studies of AHL mimics, no antagonists were uncovered in EsaR. These results provide chemical strategies to start to investigate mechanisms of ligand response in EsaR and define receptor features driving dissociative vs associative mechanisms in the LuxR-type receptor family. Our findings also suggest that alternate approaches may be required to develop competitive antagonists for dissociative LuxR-type receptors.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2451–2461"},"PeriodicalIF":3.8,"publicationDate":"2025-09-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12459020/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145079077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Characterization of Variation in Natural Product Production Under Chemical Elicitation Using Parallel Stable Isotope Labeling","authors":"Liana Zaroubi,&nbsp;, ,&nbsp;Bruno S. Paulo,&nbsp;, ,&nbsp;Ethan Fung,&nbsp;, ,&nbsp;Hannah Cavanagh,&nbsp;, ,&nbsp;Robert Britton,&nbsp;, ,&nbsp;Alessandra S. Eustaquio,&nbsp;, and ,&nbsp;Roger G. Linington*,&nbsp;","doi":"10.1021/acschembio.5c00346","DOIUrl":"10.1021/acschembio.5c00346","url":null,"abstract":"<p >Most microorganisms produce far fewer secondary metabolites under laboratory culture conditions than would be expected based on the number of biosynthetic gene clusters (BGCs) present in their genomes. One strategy for inducing secondary metabolite production is to add chemical elicitors that disrupt bacterial metabolism. This one-strain-many-compounds (OSMAC) strategy has been used successfully to discover a broad range of natural products. However, traditional strategies for detecting changes in natural product production are not well suited to characterizing variations in the full secondary metabolome under elicitation conditions. One efficient tool to differentiate metabolites between experiments is IsoAnalyst, a parallel stable isotope labeling method that connects secondary metabolites to BGCs by determining the rates of incorporation for a set of isotopically labeled secondary metabolism building blocks. In this study three strains of <i>Paraburkholderia</i> were profiled under a range of OSMAC conditions and changes in secondary metabolism characterized using a combination of analytical tools including IsoAnalyst. Using these profiles, we assessed the degree of novel secondary metabolite production under different elicitation conditions. Prioritization of one compound class strongly induced in the presence of the antibiotic rifaximin led to the discovery of 2-hydroxyacyl putrescine compounds putrescinamides A (<b>1</b>) and B (<b>2</b>). The structures of these new metabolites were determined through a combination of multidimensional NMR experiments and total synthesis, which permitted the determination of their full absolute configurations. Together these stable isotope labeling experiments provide a unique perspective on system-wide variation in de novo secondary metabolite biosynthesis under elicitor conditions and highlight the impact of elicitor selection on metabolite induction in Burkholderiales strains.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2393–2403"},"PeriodicalIF":3.8,"publicationDate":"2025-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145074045","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metabolic Tracing of Methyl Donor Utilization in Histone Methylation via Relative Quantification of Isotopomer Distribution Mass Spectrometry","authors":"Hui Tang,&nbsp; and ,&nbsp;Kangling Zhang*,&nbsp;","doi":"10.1021/acschembio.5c00528","DOIUrl":"10.1021/acschembio.5c00528","url":null,"abstract":"<p >Histone methylation depends on one-carbon metabolism, with methyl groups donated by methionine-, serine-, and glucose-derived intermediates. To dissect the metabolic origins of histone methylation, we developed Relative Quantitative Methyl Isotopomer Distribution Mass Spectrometry (RQMID-MS), a high-resolution mass spectrometry-based method that uses diagnostic low-mass fragment ions to quantify methyl group transfer from isotope-labeled precursors. Using this method, we mapped methylation sources to histone lysines in glioblastoma cells under nutrient and oxygen stress. Methionine was the dominant methyl donor under replete condition. Under combined serine and methionine depletion or prolonged methionine depletion alone, glucose emerged as a key compensatory source, particularly in U87 cells with elevated 3-phosphoglycerate dehydrogenase (PHGDH) expression. In contrast, U251 cells favored exogenous serine and glycine, correlating with higher levels of serine hydroxymethyltransferase 2 (SHMT2) expression. Hypoxia initially enhanced glucose-derived methylation but later suppressed it, likely due to impaired vitamin B₁₂-dependent remethylation of homocysteine. RQMID-MS enables precise tracking of methyl donor routing to histones and offers a robust platform for studying metabolic and epigenetic crosstalk in cancer and beyond.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2483–2493"},"PeriodicalIF":3.8,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145074003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mixed Alkyl Aryl Phosphonates as Quenched Activity-Based Probes for Real-Time Imaging of Active Neutrophil Serine Proteases","authors":"Jan Pascal Kahler,&nbsp;, ,&nbsp;Jonathan Coene,&nbsp;, ,&nbsp;Marcin Skorenski,&nbsp;, ,&nbsp;Dimitris Korovesis,&nbsp;, and ,&nbsp;Steven H. L. Verhelst*,&nbsp;","doi":"10.1021/acschembio.4c00860","DOIUrl":"10.1021/acschembio.4c00860","url":null,"abstract":"<p >Activity-based probes have been instrumental in the study of proteases, and quenched fluorescent versions can be utilized in real time imaging. Unfortunately, this application has not yet been reported for serine proteases, which make up the largest mechanistic class of proteases. Here, we describe quenched activity-based probes for detection of serine proteases, specifically the neutrophil serine proteases: neutrophil elastase, proteinase 3, and cathepsin G. We demonstrate that these reagents can selectively label serine proteases in complex proteomes and we illustrate their use in the live cell imaging of activation of primary human neutrophils. We expect that these reagents will find use in real-time imaging of active neutrophil serine proteases and may be further developed for imaging of other serine proteases.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2356–2362"},"PeriodicalIF":3.8,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145068668","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Split NeissLock with Spy-Acceleration Arms Mammalian Proteins for Anhydride-Mediated Cell Ligation","authors":"Sheryl Y. T. Lim,&nbsp;, ,&nbsp;Anthony H. Keeble,&nbsp;, and ,&nbsp;Mark R. Howarth*,&nbsp;","doi":"10.1021/acschembio.5c00515","DOIUrl":"10.1021/acschembio.5c00515","url":null,"abstract":"<p >Reactive functional groups may be incorporated into proteins or may emerge from natural amino acids in exceptional architectures. Anhydride formation is triggered by calcium in the self-processing module (SPM) of <i>Neisseria meningitidis</i> FrpC, which we previously engineered for “NeissLock” ligation to an unmodified target protein. Here, we explored bacterial diversity, discovering a related module with ultrafast anhydride formation. We dissected this swift SPM to generate a split NeissLock system, providing a second layer of control of anhydride generation: first mixing N- and C-terminal NeissLock moieties and second adding millimolar amounts of calcium. Split NeissLock generated a minimal fusion tag, permitting binder expression in mammalian cells with complex post-translational modifications and avoiding self-cleavage while transiting the calcium-rich secretory pathway. Employing spontaneous amidation between SpyTag003 and SpyCatcher003, we dramatically accelerated split NeissLock reconstitution, allowing a rapid high-yield reaction to naturally occurring targets. We established a specific covalent reaction to endogenous Epidermal Growth Factor Receptor using split NeissLock via Transforming Growth Factor-α secreted from mammalian cells. Modular ligation was demonstrated on living cells through site-specific coupling of the clot-busting enzyme tissue plasminogen activator or a computationally designed cytokine. Split NeissLock provides a modular architecture to generate highly reactive functionality, with inducibility and simple genetic encoding for enhanced cellular modification.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2475–2482"},"PeriodicalIF":3.8,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acschembio.5c00515","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145062916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Discovery and Structural Optimization of 2-Hydrazinopyrimidin-4-one Analogs Inhibiting Human ADP-Ribosylhydrolase ARH3","authors":"Tomi A.O. Parviainen,&nbsp;, ,&nbsp;Men Thi Hoai Duong,&nbsp;, ,&nbsp;Johan Pääkkönen,&nbsp;, ,&nbsp;Kamila Burdova,&nbsp;, ,&nbsp;Barbora Kuttichova,&nbsp;, ,&nbsp;Hana Hanzlikova,&nbsp;, ,&nbsp;Lari Lehtiö*,&nbsp;, and ,&nbsp;Juha P. Heiskanen*,&nbsp;","doi":"10.1021/acschembio.5c00461","DOIUrl":"10.1021/acschembio.5c00461","url":null,"abstract":"<p >Poly-ADP-ribosylation at sites of DNA damage, catalyzed by PARP enzymes, activates the DNA damage response, chromatin remodeling, and DNA repair. The modification is reversed by two enzymes in humans: PARG, which efficiently hydrolyzes the poly-ADP-ribose chains, and ARH3, which is the key enzyme for removing the last proximal mono-ADP-ribose from serine residues. While inhibitor development has largely focused on PARPs and PARG, no potent and selective inhibitors for ARH3 are currently available. We optimized a FRET-based competition assay for ARH3 and carried out high-throughput screening of small-molecule inhibitors. One hit compound, <b>1</b>, with a potency of 22 μM was discovered, and through structure–activity relationship studies and synthesis, we improved its potency 10-fold to 2 μM (compound <b>27</b>, MDOLL-0286). We demonstrate that the compound inhibits ARH3’s poly-ADP-ribose hydrolytic activity on cellular substrates. Intriguingly, it does not effectively inhibit the hydrolysis of mono-ADP-ribosylation from natural protein substrates. This is despite the fact that the cocrystal structure of compound <b>1</b> bound to ARH3 reveals its overlap with the enzyme’s ADP-ribose binding site, agreeing with the competition in the FRET assay. The first experimental ARH3 inhibitor complex provides a valuable starting point for developing more potent chemical probes to study DNA damage response mechanisms in the future.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2438–2450"},"PeriodicalIF":3.8,"publicationDate":"2025-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145062883","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Enzymatic Construction of Rare Pyrazino[1,2-a]indole Framework: Side Chain Migration-Driven Pictet–Spengler Activity of McbB","authors":"Haicheng Liu,&nbsp;, ,&nbsp;Wangtao Jiang,&nbsp;, ,&nbsp;Jinbiao Li,&nbsp;, ,&nbsp;Yushi Futamura,&nbsp;, ,&nbsp;Hiroyuki Osada,&nbsp;, and ,&nbsp;Hongbin Zou*,&nbsp;","doi":"10.1021/acschembio.5c00455","DOIUrl":"10.1021/acschembio.5c00455","url":null,"abstract":"<p >Rational substrate exploration with enzymes can unlock innovative molecular architectures and reveal unprecedented biological activities. Inspired by our previous studies on strictosidine synthase, a plant-derived Pictet–Spenglerase, we designed (<i>S</i>)-2-amino-3-(1<i>H</i>-indol-1-yl)propionic acid (<b>1</b>) as a novel substrate for the microbial Pictet–Spenglerase McbB. Enzymatic condensation of <b>1</b> with oxaloacetaldehyde afforded the rare pyrazino[1,2-<i>a</i>]indole scaffold, overcoming both intrinsic β-carboline bias and the limitations of conventional chemical Pictet–Spengler reactions. Substrate profiling revealed that McbB exhibited strict specificity for 4-methyl- and 5-fluoro-substituted analogs of <b>1</b>, yet tolerated methylglyoxal or formaldehyde as aldehyde partners with <b>1</b>. Site-directed mutagenesis combined with computational docking delineated a new substrate binding mode and provided insights into the catalytic mechanism. Further chemoenzymatic derivatization of 1 yielded novel pyrazino[1,2-<i>a</i>]indoles with notable antiplasmodial (compound <b>12</b>, IC₅₀ = 1.5 ± 0.2 μM vs <i>P.f.</i> 3D7) and antitumor activities (compound <b>13</b>, IC₅₀ = 3.19 ± 0.3 μM vs HL60). This study expands our understanding of the enzymatic mechanism of McbB and enables the development of diverse bioactive compounds through substrate exploration.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2428–2437"},"PeriodicalIF":3.8,"publicationDate":"2025-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145051397","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis and Antimicrobial Specificities of Halogenated Tryptophan-Containing Nisin Variants","authors":"Chenhui Wang,&nbsp;, ,&nbsp;Sanne Tervoort,&nbsp;, ,&nbsp;Oscar P. Kuipers,&nbsp;, and ,&nbsp;Jaap Broos*,&nbsp;","doi":"10.1021/acschembio.5c00632","DOIUrl":"10.1021/acschembio.5c00632","url":null,"abstract":"<p >Antimicrobial peptides, and in particular ribosomally produced and post-translationally modified peptides (RiPPs), are a potentially important class of candidate antibiotics for combating multidrug-resistant bacteria. Introduction of a halogenated Trp residue into a RiPP can possibly enhance antimicrobial efficacy and alter specificity, but this modification has hardly been explored. This study employs an efficient expression system utilizing a tryptophan auxotrophic <i>Lactococcus lactis</i> strain to biosynthetically and efficiently incorporate halogenated tryptophan analogues, namely 5-fluoro-tryptophan (5FW), 5-chloro-tryptophan (5CW), 5-bromo-tryptophan (5BW), as well as 5-methyl-tryptophan (5MW) at position 1 of I1W nisin A. Wild-type nisin and Trp-containing I1W nisin show a high and broad activity against four tested pathogens. However, the activity spectrum of the three different halogen atom containing nisin variants became more strain specific, as both increased and decreased activities were measured against the four tested pathogens. No trend between the chemical properties of the halogen atom (e.g., electronegativity, size) and the bioactivity of the nisin variants toward each of the four pathogens could be detected, suggesting strain specific antimicrobial activity mechanisms. These findings demonstrate that halogenated tryptophan analogues can be successfully incorporated into a bioactive RiPP produced by an auxotrophic <i>L. lactis</i> strain and underscore the utility of peptide halogenation for discovering novel antimicrobial agents with tailored pathogen specificity.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 10","pages":"2503–2511"},"PeriodicalIF":3.8,"publicationDate":"2025-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/pdf/10.1021/acschembio.5c00632","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145051387","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}