ACS Chemical Biology最新文献

Targeted O-GlcNAcylation of CK2α Triggers Its Ubiquitin-Proteasome Degradation and Alters Downstream Phosphorylation. CK2α靶向o - glcn酰化触发其泛素蛋白酶体降解并改变下游磷酸化。

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-16 DOI: 10.1021/acschembio.5c00223

Tongyang Xu, Bowen Ma, Yuanpei Li, Zhihao Guo, Miaomiao Zhang, Billy Wai-Lung Ng

{"title":"Targeted O-GlcNAcylation of CK2α Triggers Its Ubiquitin-Proteasome Degradation and Alters Downstream Phosphorylation.","authors":"Tongyang Xu, Bowen Ma, Yuanpei Li, Zhihao Guo, Miaomiao Zhang, Billy Wai-Lung Ng","doi":"10.1021/acschembio.5c00223","DOIUrl":"https://doi.org/10.1021/acschembio.5c00223","url":null,"abstract":"O-Linked β-N-acetylglucosamine-modification (O-GlcNAcylation) is an important post-translational modification (PTM), yet dissecting its protein-specific functions has remained challenging. Here, we applied our previously reported chemical biology tool, the O-GlcNAcylation Targeting Chimera (OGTAC), to specifically induce O-GlcNAcylation of the casein kinase II subunit α (CK2α) at Ser347 in living cells. We found that this targeted O-GlcNAcylation destabilized CK2α through ubiquitin-proteasome degradation and enhanced its interaction with cereblon (CRBN). Overexpression and knockdown experiments also indicated CK2α as a substrate of the Cullin-RING E3 ubiquitin ligase 4-CRBN (CRL4CRBN) E3 ligase complex. Furthermore, the OGTAC-induced O-GlcNAcylation of CK2α reprogrammed phosphorylation of Akt and PFKP. These findings reveal that a single O-GlcNAc modification can serve as a molecular switch, controlling the protein stability and downstream phosphorylation of CK2α. More broadly, our results highlight the profound utility of the OGTAC-mediated O-GlcNAcylation to interrogate its cellular functions with specificity, overcoming limitations inherent to prior global perturbation methods.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144300535","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}

引用次数: 0

Investigation of Glutarimide N-Alkylated Derivatives of Lenalidomide. 来那度胺戊二胺n -烷基化衍生物的研究。

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-16 DOI: 10.1021/acschembio.5c00272

Farah Kabir, Toshiaki Sonobe, Qian Zhu, Nandini Vallavoju, Yuka Amako, Christina M Woo

{"title":"Investigation of Glutarimide N-Alkylated Derivatives of Lenalidomide.","authors":"Farah Kabir, Toshiaki Sonobe, Qian Zhu, Nandini Vallavoju, Yuka Amako, Christina M Woo","doi":"10.1021/acschembio.5c00272","DOIUrl":"https://doi.org/10.1021/acschembio.5c00272","url":null,"abstract":"Lenalidomide is a thalidomide derivative that engages the E3 ligase substrate receptor cereblon (CRBN) to promote targeted protein degradation. Lenalidomide possesses a glutarimide moiety, which is responsible for CRBN engagement, and an isoindoline moiety, which promotes neosubstrate recruitment. Modification of the glutarimide is a generalizable prodrug strategy to inhibit CRBN binding for the selective activation of CRBN-dependent activity, yet these compounds may possess CRBN-independent effects. We prepared six N-alkylated glutarimide derivatives and found CRBN-independent effects on TNFα inhibition and selective effects in the cell viability profiles. Evaluation of selected compounds by global proteomics in KG1a cells reveals that the downregulation of Rab28 is CRBN-independent and mediated by autophagy. Finally, we developed a representative prodrug to demonstrate the enzymatic release of lenalidomide. Collectively, although some CRBN-independent properties are observed, modification of glutarimide is a generally viable strategy to prevent CRBN engagement in a prodrug strategy.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144309239","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}

引用次数: 0

Biphasic Cellular Response Triggered by Tetrandrine-Mediated Dysfunction and Lysophagic Clearance of Lysosomes. 由粉防己碱介导的功能障碍和溶酶体的溶噬清除引发的双相细胞反应。

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-16 DOI: 10.1021/acschembio.5c00220

Zhe Yang, Tomoki Takahashi, Ayase Hoshino, Tatsuya Yamamoto, Hideyuki Shigemori, Yusaku Miyamae

{"title":"Biphasic Cellular Response Triggered by Tetrandrine-Mediated Dysfunction and Lysophagic Clearance of Lysosomes.","authors":"Zhe Yang, Tomoki Takahashi, Ayase Hoshino, Tatsuya Yamamoto, Hideyuki Shigemori, Yusaku Miyamae","doi":"10.1021/acschembio.5c00220","DOIUrl":"10.1021/acschembio.5c00220","url":null,"abstract":"Lysosomes play an important role in the degradation of cellular components and are correlated with various other physiological phenomena. Lysophagy is a cellular quality control system that maintains homeostasis by removing damaged lysosomes through autophagy. The involvement of lysosomal dysfunction in the pathogenesis of certain illnesses (e.g., neurodegeneration) highlights the potential of small molecules that regulate lysophagy as drug candidates. Here, we found that tetrandrine, a bis-benzylisoquinoline alkaloid, induces lysophagy, leading to the clearance of damaged lysosomes in mammalian cells. To visualize the target organelles of tetrandrine, we synthesized a chimeric compound in which tetrandrine was connected to boron-dipyrromethene via a polyethylene glycol linker. Flow cytometry analysis confirmed the cellular uptake of the synthesized probe. An organelle-staining assay showed that the fluorescent signal of the probe was specifically colocalized with lysosomes. Tetrandrine transiently increased the lysosomal pH level, which returned to normal at 24 h post treatment. Consistently, the level of mCherry-tagged galectin-3, a marker protein for lysophagy, transiently increased and then diminished under treatment with tetrandrine. Tetrandrine also induced dephosphorylation of transcription factor EB, a regulator of lysosomal biogenesis, promoting its translocation from the cytosol to the nucleus. These results suggest that tetrandrine induces a biphasic cellular response, first disrupting lysosomal function before facilitating cellular lysosomal homeostasis through lysophagy and lysosomal biogenesis. This dual effect distinguishes tetrandrine from existing lysosomal modulators.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144300533","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}

引用次数: 0

Characterization of the Postaglycone Modifications in Ristomycin Biosynthesis. 利斯托霉素生物合成中后苷元修饰的表征。

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-15 DOI: 10.1021/acschembio.5c00280

Zhanzhao Cui, Xiaozheng Wang, Lixin Yin, Lu Chen, Kai Liu, Yemin Wang, Shuangjun Lin, Zixin Deng, Min Xu, Meifeng Tao

{"title":"Characterization of the Postaglycone Modifications in Ristomycin Biosynthesis.","authors":"Zhanzhao Cui, Xiaozheng Wang, Lixin Yin, Lu Chen, Kai Liu, Yemin Wang, Shuangjun Lin, Zixin Deng, Min Xu, Meifeng Tao","doi":"10.1021/acschembio.5c00280","DOIUrl":"https://doi.org/10.1021/acschembio.5c00280","url":null,"abstract":"Amycolatopsis sp. TNS106 produces ristomycin (ristocetin), a type III glycopeptide antibiotic (GPA) featuring extensive glycosyl modifications and potent antimicrobial activity against Gram-positive pathogens. Unlike the well-documented nonribosomal peptide synthetase-assembled peptide scaffold, the timing and specificity of its postassembly tailoring steps remain poorly understood. In this study, we generated a series of A. sp. TNS106 mutants and characterized accumulated derivatives to delineate the postaglycone tailoring steps for ristomycin maturation. In vitro biochemical reactions confirmed the function and timing of the RgtfB and RgtfC glycosyltransferases and MtfA carboxyl methyltransferase. By integrating our findings with prior studies, we propose a comprehensive model for the sequential glycosylation and C-terminal methylation processes governing ristomycin biosynthesis. Notably, we identified multiple ristomycin glycoforms that demonstrate improved antimicrobial activity compared to the parent molecule. Strikingly, removal of the ristosamine on β-hydroxytyrosine 6 (βht6) or the presence of rhamnose on 4-hydroxyphenylglycine 4 (Hpg4) markedly impaired its antibacterial activity, particularly against vancomycin-resistant Enterococcus. The engineered strains constructed here provide a versatile platform for generating novel ristomycin analogs through combinatorial biosynthesis or chemical synthesis, advancing the development of new-to-nature GPAs with improved potency.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144300534","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}

引用次数: 0

Discovery of Inhibitors for Bacterial Arr Enzymes ADP-Ribosylating and Inactivating Rifamycin Antibiotics. 细菌Arr酶adp -核糖基化和利福霉素抗生素灭活抑制剂的发现。

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-13 DOI: 10.1021/acschembio.5c00164

Juho Alaviuhkola, Sondos Abdulmajeed, Sven T Sowa, Johan Pääkkönen, Lari Lehtiö

{"title":"Discovery of Inhibitors for Bacterial Arr Enzymes ADP-Ribosylating and Inactivating Rifamycin Antibiotics.","authors":"Juho Alaviuhkola, Sondos Abdulmajeed, Sven T Sowa, Johan Pääkkönen, Lari Lehtiö","doi":"10.1021/acschembio.5c00164","DOIUrl":"https://doi.org/10.1021/acschembio.5c00164","url":null,"abstract":"ADP-ribosylation is an enzymatic process where an ADP-ribose moiety is transferred from NAD+ to an acceptor molecule. While ADP-ribosylation is well-established as a post-translational modification of proteins, rifamycin antibiotics are its only known small-molecule targets. ADP-ribosylation of rifampicin was first identified in Mycolicibacterium smegmatis, whose Arr enzyme transfers the ADP-ribose moiety to the 23-hydroxy group of rifampicin preventing its interaction with the bacterial RNA polymerase thereby inactivating the antibiotic. Arr homologues are widely spread among bacterial species and present in several pathogenic species often associated with mobile genetic elements. Inhibition of Arr enzymes offers a promising strategy to overcome ADP-ribosylation mediated rifamycin resistance. We developed a high-throughput activity assay which was applied to screen an in-house library of human ADP-ribosyltransferase-targeted compounds. We identified 15 inhibitors with IC50 values below 5 μM against four Arr enzymes from M. smegmatis, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, and Mycobacteroides abscessus. The observed overall selectivity of the hit compounds over the other homologues indicated structural differences between the proteins. We crystallized M. smegmatis and P. aeruginosa Arr enzymes, the former in complex with its most potent hit compound with an IC50 value of 1.3 μM. We observed structural differences in the NAD+ binding pockets of the two Arr homologues explaining the selectivity. Although the Arr inhibitors did not sensitize M. smegmatis to rifampicin in a growth inhibition assay, the structural information and the collection of inhibitors provide a foundation for rational modifications and further development of the compounds.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144281662","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}

引用次数: 0

Functional Conservation and Divergence of AlpJ-Family Oxygenases Catalyzing C-C Bond Cleavage in Atypical Angucycline Biosynthesis. 非典型安环素合成中催化C-C键断裂的alpj家族加氧酶的功能保守与分化

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-11 DOI: 10.1021/acschembio.5c00040

Shijie Shen, Changbiao Chi, Keqiang Fan, Qian Zhang, Yang Xu, Jinmin Gao, Huitao Hu, Lijun Wang, Donghui Yang, Ming Ma, Guohui Pan

{"title":"Functional Conservation and Divergence of AlpJ-Family Oxygenases Catalyzing C-C Bond Cleavage in Atypical Angucycline Biosynthesis.","authors":"Shijie Shen, Changbiao Chi, Keqiang Fan, Qian Zhang, Yang Xu, Jinmin Gao, Huitao Hu, Lijun Wang, Donghui Yang, Ming Ma, Guohui Pan","doi":"10.1021/acschembio.5c00040","DOIUrl":"https://doi.org/10.1021/acschembio.5c00040","url":null,"abstract":"AlpJ-family oxygenases catalyze distinctive oxidative B-ring cleavage and rearrangement reactions during the biosynthesis of atypical angucycline natural products, which are characterized by unique chemical structures and diverse biological activities. While the individual functions of a few AlpJ-family enzymes have been reported, there is a lack of systematic exploration and functional comparison within this enzyme family, hindering a comprehensive understanding of the AlpJ-family oxygenases. In this study, we have systematically explored and analyzed AlpJ-family oxygenases, identifying 49 representative homologues, which can be classified into two distinct evolutionary groups. We revealed that enzymes from different groups exhibit clear functional differentiation, catalyzing the same angucycline substrate dehydrorabelomycin into distinct products, whereas enzymes within the same group display more similar catalytic functions with varying degrees of functional overlap. This underscores the intriguing functional conservation and divergence of the AlpJ-family oxygenases. In addition, we report the first crystal structure of a Group I enzyme, PenE. Structural analysis and site-directed mutagenesis identified key structural features and residues within AlpJ-family oxygenases, which harbor hydrophobic substrate-binding pockets at both the N- and C-termini, both of which are essential for function. Our findings provide valuable insights into the evolution, catalytic mechanisms, and functional divergence of this unique family of oxygenases. Further investigation of these newly identified AlpJ homologues and their associated biosynthetic gene clusters will facilitate the discovery of enzymes with unique catalytic mechanisms and bioactive atypical angucyclines with novel structures.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144273649","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}

引用次数: 0

High-Throughput Screening Tool to Identify Small Molecule Inhibitors of Telomerase. 鉴别端粒酶小分子抑制剂的高通量筛选工具。

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-10 DOI: 10.1021/acschembio.5c00244

Elisa Aquilanti, Sulyman Barkho, Vincent Bozinov, Lauren Kageler, Max Garrity-Janger, Michael F Mesleh, Steven Horner, Matthew J Ranaghan, Matthew Meyerson

{"title":"High-Throughput Screening Tool to Identify Small Molecule Inhibitors of Telomerase.","authors":"Elisa Aquilanti, Sulyman Barkho, Vincent Bozinov, Lauren Kageler, Max Garrity-Janger, Michael F Mesleh, Steven Horner, Matthew J Ranaghan, Matthew Meyerson","doi":"10.1021/acschembio.5c00244","DOIUrl":"10.1021/acschembio.5c00244","url":null,"abstract":"Telomerase reverse transcriptase is a ribonucleoprotein complex that maintains telomere length in rapidly dividing cells, thus enabling cellular immortality. Despite being recognized as an important cancer target for decades, no small molecule telomerase inhibitors have been approved as anticancer therapeutics to date. Several limitations, including the absence of high-throughput screening tools, have posed challenges to the telomerase drug discovery field. Here, we describe a high-throughput, fluorescently coupled screening method employing a chemically modified reporter nucleotide. We utilize the Tribolium castaneum telomerase as a surrogate model as it shares a high degree of active site homology with the human enzyme. We piloted this tool by screening a chemical library of ∼3600 nucleoside mimetics to demonstrate excellent assay quality, and identified 2 compounds with inhibitory activity that were further validated in a direct enzymatic assay. Our work introduces a method that has the potential to uncover novel telomerase inhibitors for further drug discovery efforts.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144256626","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}

引用次数: 0

Experimental and Computational Evaluation of Nicotinamide Cofactor Biomimetics. 烟酰胺辅助因子仿生的实验与计算评价。

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-09 DOI: 10.1021/acschembio.5c00174

Karissa C Kenney, Tyler P LaFortune, Sourav Majumdar, Edgar M Manriquez, Arjun S Pamidi, Courtnie S Kom, Jason E Garrido, Edgar S Villa, Filipp Furche, Gregory A Weiss

{"title":"Experimental and Computational Evaluation of Nicotinamide Cofactor Biomimetics.","authors":"Karissa C Kenney, Tyler P LaFortune, Sourav Majumdar, Edgar M Manriquez, Arjun S Pamidi, Courtnie S Kom, Jason E Garrido, Edgar S Villa, Filipp Furche, Gregory A Weiss","doi":"10.1021/acschembio.5c00174","DOIUrl":"https://doi.org/10.1021/acschembio.5c00174","url":null,"abstract":"Oxidoreductase enzymes are widely used biocatalysts due to their high enantioselectivity and broad substrate compatibility in useful transformations. Many oxidoreductases require nicotinamide cofactors (i.e., NAD(P)H). To replace this costly natural cofactor, synthetic nicotinamide cofactor biomimetics (NCBs) offer different shapes, binding affinities, and reducing potentials that exceed the capabilities of wild-type NAD(P)H. However, the ill-defined structure-activity relationships (SARs) of various NCBs slow rationally guided innovation, such as customized reducing potentials. Here, we dissect two essential elements of NCB design, holding the nicotinamide invariant. First, the linker length between the nicotinamide and an unconjugated aromatic ring uncovered unexpected benefits to redox activity for two or three carbon linkers. Second, substitution on this unconjugated aryl group (Ring 2) might not be expected to affect activity. However, SAR trends demonstrate substantial benefits to reductive potential conferred by electron-donating functionalities on Ring 2. Furthermore, catalysis by two enzymes demonstrates enzyme-dependent tolerance or sensitivity to the NCB structures. Density functional theory (DFT) and computational modeling provide a theoretical framework to understand and build upon these observations. Ring 2 reaches up to the nicotinamide to stabilize its positive charge after oxidation through π-π stacking and charge transfer. Thus, the systematic examination of NCB's stability, electrochemical redox potentials, and kinetics uncovers trends for the improved design of NCBs.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144256625","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}

引用次数: 0

DNA Alkylation, Cross-Linking, and Cancer Cell Killing by a Quinoxaline-N-Mustard Conjugate. DNA烷基化，交联和癌细胞杀伤喹啉- n -芥菜缀合物。

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-06 DOI: 10.1021/acschembio.4c00734

Chandra Sova Mandi, Dipendu Patra, Tanhaul Islam, Bhim Majhi, Kent S Gates, Sanjay Dutta

{"title":"DNA Alkylation, Cross-Linking, and Cancer Cell Killing by a Quinoxaline-N-Mustard Conjugate.","authors":"Chandra Sova Mandi, Dipendu Patra, Tanhaul Islam, Bhim Majhi, Kent S Gates, Sanjay Dutta","doi":"10.1021/acschembio.4c00734","DOIUrl":"https://doi.org/10.1021/acschembio.4c00734","url":null,"abstract":"Nitrogen mustards are a family of clinically used anticancer drugs that contain a DNA-alkylating bis(2-chloroethyl)amino group. Appending the bis(2-chloroethyl)amino alkylating agent to noncovalent DNA-binding groups such as intercalators, polyamides, or polyamines has the potential to yield DNA-targeted anticancer agents with improved potency. In the work reported here, substituted quinoxaline groups were explored as minimal intercalators expected to confer noncovalent DNA-binding properties on a bis(2-chloroethyl)anilino mustard alkylating unit. A quinoxaline unit with a cationic dimethylamino-containing side chain was found to be a more potent DNA-alkylating and cross-linking agent than the clinically used mustard chlorambucil (Chb). The results of dye displacement and multiple DNA alkylation assays showed that the quinoxaline ring binds noncovalently to duplex DNA, likely via intercalation. The quinoxaline-mustard conjugate was more active than Chb against a variety of cancer cell lines. Evidence is presented, showing that both the quinoxaline-mustard and the clinically used drug Chb formed aggregates in aqueous buffer; however, the results clearly show that the propensity to form aggregates clearly does not abrogate the DNA-alkylating properties or bioactivity of these compounds.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144245188","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}

引用次数: 0

Identification and Characterization of the Biosynthesis of the Hybrid NRPS-NIS Siderophore Nocardichelin. nrpps - nis铁载体诺卡地helin的生物合成鉴定与表征。

IF 3.5 2区生物学

ACS Chemical Biology Pub Date : 2025-06-06 DOI: 10.1021/acschembio.5c00286

Mercedes B Fisk, Jocelyn Barrera Ramirez, Collin E Merrick, Timothy A Wencewicz, Andrew M Gulick

{"title":"Identification and Characterization of the Biosynthesis of the Hybrid NRPS-NIS Siderophore Nocardichelin.","authors":"Mercedes B Fisk, Jocelyn Barrera Ramirez, Collin E Merrick, Timothy A Wencewicz, Andrew M Gulick","doi":"10.1021/acschembio.5c00286","DOIUrl":"https://doi.org/10.1021/acschembio.5c00286","url":null,"abstract":"Bacteria cope with the limitation of iron by producing siderophores, small molecules they export that have high affinity for iron. Once complexed, the ferric siderophore is transported into the cell through specialized receptors allowing the iron to be released and used in a variety of biological processes. Many peptide siderophores that use catechol, phenolate, or oxazoline/thiazoline groups to coordinate iron are produced by a family of enzymes called nonribosomal peptide synthetases (NRPSs). Alternately, a smaller family of NRPS-independent siderophores (NISs) is produced by a different biosynthetic strategy. The NIS pathways employ one or more NIS synthetases that combine an amine commonly harboring a hydroxamate with a carboxylate substrate. Discovered in 2007 in an uncharacterized Nocardia species, a siderophore called nocardichelin was identified and chemically characterized that contained features of both NIS and NRPS siderophores. Nocardichelin contains an N-salicyloxazoline moiety, predicted to be built by a modular NRPS, and a dihydroxamate containing N-hydroxy-N-succinylcadaverine and N-hydroxy-N-tetradecenoylcadaverine groups. To explore this potential hybrid NRPS/NIS, we identified a biosynthetic gene cluster in Nocardia carnea containing 13 enzymes and four proteins involved in transport. We have functionally characterized four of the enzymes for their activity and substrate specificity and further solved the structures of two enzymes. We present our discovery and initial characterization of this cluster, describe remaining questions for elucidation of the unusual siderophore, and discuss the potential for use in downstream biocatalytic applications.","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-06-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144245189","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}

引用次数: 0