ACS Chemical Biology最新文献

{"title":"Optimizing Peptide-Conjugated Lipid Nanoparticles for Efficient siRNA Delivery across the Blood-Brain Barrier and Treatment of Glioblastoma Multiforme.","authors":"Haiyang Tong, Zesen Ma, Jin Yu, Dongsheng Li, Qingjun Zhu, Huajian Shi, Yun Wu, Hongyi Yang, Yanmin Zheng, Demeng Sun, Pan Shi, Jiaru Chu, Pei Lv, Baoqing Li, Changlin Tian","doi":"10.1021/acschembio.5c00039","DOIUrl":"https://doi.org/10.1021/acschembio.5c00039","url":null,"abstract":"<p><p>Glioblastoma multiforme (GBM) is a WHO grade 4 glioma and the most common malignant primary brain tumor. Addressing the clinical management of GBM presents an exceptionally daunting and intricate challenge, particularly in overcoming the blood-brain barrier (BBB) to deliver effective therapies to the brain. Nanotechnology-based drug delivery systems have exhibited considerable promise in tackling this aggressive brain cancer. However, the BBB remains a key challenge in achieving effective brain delivery of nanocarriers. Here, we have optimized a lipid nanoparticle (LNP) formulation (C2) and modified the LNP with Angiopep-2 peptide, which exhibits the most significant improvements in blood-brain barrier penetration and brain accumulation (about 2.23% injection dose). Using the Ang-2-coupled C2 LNP formulation, we researched the therapeutic effect of Polo-like Kinase 1(PLK1)-targeted siRNA delivery to treat a mouse model of GBM. The optimized LNP formulation was demonstrated to significantly inhibit mouse GBM growth and extend the median survival of mice (2.18-fold). This work demonstrates the efficacy of a brain-targeted siRNA delivery system in GBM treatment. As the understanding of the role of RNAs in GBM deepens and innovative delivery methods are continually developed and refined, RNA-based therapies could emerge as a crucial breakthrough in the advancement of brain tumor treatment.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143622811","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":"In Situ Labeling of Pathogenic Tau Using Photo-Affinity Chemical Probes","authors":"Pengju Nie,&nbsp;You Wu*,&nbsp;John Robinson,&nbsp;Shekar Mekala,&nbsp;Virginia M. Y. Lee and Yue-Ming Li*,&nbsp;","doi":"10.1021/acschembio.5c0007310.1021/acschembio.5c00073","DOIUrl":"https://doi.org/10.1021/acschembio.5c00073https://doi.org/10.1021/acschembio.5c00073","url":null,"abstract":"<p >Tau aggregation plays a crucial role in the development of Alzheimer’s disease (AD). Developing specific techniques that can isolate pathogenic tau from brain tissue is important for understanding tauopathies and advancing targeted therapies. Here, we develop photoaffinity small molecular probes and a novel method for <i>in situ</i> tissue labeling and investigate their activity in interacting with tau in cells and AD patient brains. Based on the reported chemical structures of tau PET tracers, we designed and synthesized two tau-specific probes, namely, Tau-2 and Tau-4. After validation in cell, mouse model, and patient brain samples, our photolabeling results suggested that Tau-2 effectively labels soluble tau in cell and mouse models, while Tau-4 selectively binds high-molecular-weight tau aggregates in late-stage AD patient brain tissues. Proteomic analysis verified the specific isolation of pathogenic tau from AD brain samples. Collectively, these findings underscore the potential of our photoaffinity probes as powerful tools for investigating tau proteins and neurofibrillary tangles in neurodegenerative diseases.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 3","pages":"581–591 581–591"},"PeriodicalIF":3.5,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143666930","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":"Rational Design of Stapled Covalent Peptide Modifiers of Oncoprotein E6 from Human Papillomavirus","authors":"Cole Emanuelson,&nbsp;Yuta Naro,&nbsp;Olivia Shade,&nbsp;Melinda Liu,&nbsp;Sagar D. Khare and Alexander Deiters*,&nbsp;","doi":"10.1021/acschembio.4c0087810.1021/acschembio.4c00878","DOIUrl":"https://doi.org/10.1021/acschembio.4c00878https://doi.org/10.1021/acschembio.4c00878","url":null,"abstract":"<p >Human Papillomavirus (HPV) is linked to multiple cancers, most significantly cervical cancer, for which HPV infection is associated with nearly all cases. Essential to the oncogenesis of HPV is the function of the viral protein E6 and its role in degrading the cell cycle regulator p53. Degradation of p53, and the resultant loss of cell cycle control, is mediated by E6 recruitment of the E3 ubiquitin ligase E6AP and subsequent ubiquitination of p53. Here, we report the design of a stapled peptide that mimics the LxxLL α-helical domain of E6AP to bind and covalently label a cysteine residue specific to HPV-16 E6. Several acrylamide- and haloacetamide-based warheads were evaluated for reactivity and specificity, and a panel of hydrocarbon-stapled peptides was evaluated for enhanced binding affinity and increased proteolytic stability. Structure-based modeling was used to rationalize the observed trends in the reactivity of the warheads and the impact of the hydrocarbon staple position on the binding affinity of the stapled peptides. The development of a proteolytically stable and reactive peptide represents a new class of peptide-based inhibitors of protein–protein interactions with a potential therapeutic value toward HPV-derived cancers.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 3","pages":"746–757 746–757"},"PeriodicalIF":3.5,"publicationDate":"2025-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acschembio.4c00878","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143666920","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":"Biochemical Characterization of Disease-Associated Variants of Human Ornithine Transcarbamylase.","authors":"Emily Micheloni, Samantha S Watson, Penny J Beuning, Mary Jo Ondrechen","doi":"10.1021/acschembio.5c00043","DOIUrl":"https://doi.org/10.1021/acschembio.5c00043","url":null,"abstract":"<p><p>Human ornithine transcarbamylase deficiency (OTCD) is the most common ureagenesis disorder in the world. OTCD is an X-linked genetic deficiency in which patients experience hyperammonemia to varying degrees depending on the severity of the genetic mutation. More than two-thirds of the known mutations are caused by single nucleotide substitutions. In this paper, partial order optimum likelihood (POOL), a machine learning method, is used to analyze single nucleotide substitutions in OTC with varying disease phenotypes and predicted catalytic efficiencies. Specifically, we used a computed metric, μ₄, a measure of the degree of coupling between an ionizable residue and its neighbors, calculated for the catalytic residues, to identify which protein variants were most likely to have impacted catalytic activities. From this analysis, 17 disease-associated variants were selected plus one additional variant, representing a range of μ₄ values and POOL ranks. Then μ₄ predictions were compared with established bioinformatics tools, SIFT, PolyPhen-2, Provean, FATHMM, MutPred2, and MutationTaster2. The bioinformatics tools predicted that most of these mutations are deleterious. The variants were biochemically characterized using kinetics assays, size exclusion chromatography, and differential scanning fluorimetry. POOL combined with μ₄ analysis was able to predict correctly which variants were catalytically hindered in vitro for 17 out of 18 variants. Then by expressing a subset of these proteins in cell culture, mechanisms for disease were proposed. Analysis using μ₄ is a complementary method to the sequence-based bioinformatics tools for predicting the effects of mutation on catalytic function.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-03-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143583756","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":"Advances in Optogenetics and Thermogenetics for Control of Non-Neuronal Cells and Tissues in Biomedical Research","authors":"Vera S. Ovechkina*,&nbsp;Sofya K. Andrianova,&nbsp;Iana O. Shimanskaia,&nbsp;Polina S. Suvorova,&nbsp;Anna Y. Ryabinina,&nbsp;Mikhail L. Blagonravov,&nbsp;Vsevolod V. Belousov and Andrey A. Mozhaev,&nbsp;","doi":"10.1021/acschembio.4c0084210.1021/acschembio.4c00842","DOIUrl":"https://doi.org/10.1021/acschembio.4c00842https://doi.org/10.1021/acschembio.4c00842","url":null,"abstract":"<p >Optogenetics and chemogenetics are relatively new biomedical technologies that emerged 20 years ago and have been evolving rapidly since then. This has been made possible by the combined use of genetic engineering, optics, and electrophysiology. With the development of optogenetics and thermogenetics, the molecular tools for cellular control are continuously being optimized, studied, and modified, expanding both their applications and their biomedical uses. The most notable changes have occurred in the basic life sciences, especially in neurobiology and the activation of neurons to control behavior. Currently, these methods of activation have gone far beyond neurobiology and are being used in cardiovascular research, for potential cancer therapy, to control metabolism, etc. In this review, we provide brief information on the types of molecular tools for optogenetic and thermogenetic methods─microbial rhodopsins and proteins of the TRP superfamily─and also consider their applications in the field of activation of non-neuronal tissues and mammalian cells. We also consider the potential of these technologies and the prospects for the use of optogenetics and thermogenetics in biomedical research.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 3","pages":"553–572 553–572"},"PeriodicalIF":3.5,"publicationDate":"2025-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143667069","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":"Structural and Mechanistic Characterization of the Flavin-Dependent Monooxygenase and Oxidase Involved in Sorbicillinoid Biosynthesis","authors":"Gwen Tjallinks,&nbsp;Nicolò Angeleri,&nbsp;Quoc-Thai Nguyen,&nbsp;Barbara Mannucci,&nbsp;Mark Arentshorst,&nbsp;Jaap Visser,&nbsp;Arthur F. J. Ram,&nbsp;Marco W. Fraaije* and Andrea Mattevi*,&nbsp;","doi":"10.1021/acschembio.4c0078310.1021/acschembio.4c00783","DOIUrl":"https://doi.org/10.1021/acschembio.4c00783https://doi.org/10.1021/acschembio.4c00783","url":null,"abstract":"<p >Sorbicillinoids are yellow secondary metabolites synthesized through an elegant combination of enzymatic and spontaneous biochemical processes. The flavin-dependent monooxygenase SorC and oxidase SorD are crucial in this interplay, enabling the generation of a diverse array of functionally complex sorbicillinoids. By solving the crystal structures of SorC and SorD from <i>Penicillium chrysogenum</i> with sorbicillin bound in the active site, we describe the catalytically active binding conformations, crucial for attaining enantioselective and stereoselective control in these enzymatic reactions. The structure of SorC was resolved with the cofactor FAD in its <i>out</i> state, which allowed us to identify key residues that modulate flavin mobility and other conformational changes. Catalytic residues of SorC were also confirmed by detailed characterization of wild-type and several SorC variants. Meanwhile, using a CRISPR/Cas9-based multicopy-genome integration system, we could heterologously express the flavin-dependent oxidase SorD from <i>P. chrysogenum</i> in <i>Aspergillus niger</i> with high yields and purity. This allowed us to obtain the crystal structure of SorD with sorbicillin bound in a viable catalytic conformation. Structural analysis of the obtained complex provided insights into the substrate binding pose and highlighted potentially critical active site residues. Ultimately, having both SorC and SorD at our disposal enabled us to investigate their functions and interplays in the biosynthesis of a vast array of functionally complex sorbicillinoids.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 3","pages":"646–655 646–655"},"PeriodicalIF":3.5,"publicationDate":"2025-03-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acschembio.4c00783","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143666909","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":"Toyaburgine, a Synthetic <i>N</i>-Biphenyl-Dihydroisoquinoline Inspired by Related <i>N</i>,<i>C</i>-Coupled Naphthylisoquinoline Alkaloids, with High <i>In Vivo</i> Efficacy in Preclinical Pancreatic Cancer Models.","authors":"Suresh Awale, Juthamart Maneenet, Nguyen Duy Phan, Hung Hong Nguyen, Tsutomu Fujii, Heiko Ihmels, Denisa Soost, Nasir Tajuddeen, Doris Feineis, Gerhard Bringmann","doi":"10.1021/acschembio.4c00870","DOIUrl":"10.1021/acschembio.4c00870","url":null,"abstract":"<p><p>Pancreatic cancer is a highly aggressive and lethal malignancy, with a 5-year survival rate below 10%. Traditional chemotherapy, including gemcitabine, has limited efficacy due to chemoresistance and a unique tumor microenvironment characterized by hypovascularity and nutrient deprivation. This study reports on the discovery of a new <i>N</i>-biphenyl-dihydroisoquinoline, named toyaburgine (<b>4</b>), inspired by naturally occurring <i>N,C</i>-coupled naphthylisoquinoline alkaloids. Developed through systematic structural optimization, toyaburgine is a potent anticancer agent, showing promise for pancreatic cancer treatment. It exhibits strong antiausterity activity with low nanomolar PC₅₀ values, effectively inhibiting pancreatic cancer cell viability under nutrient-deprived conditions. <i>In vitro</i>, <b>4</b> causes significant morphological changes and cancer cell death in MIA PaCa-2 cells while also inhibiting cell migration and colony formation, which indicates its antimetastatic potential. Mechanistically, toyaburgine disrupts the PI3K/Akt/mTOR pathway, essential for pancreatic cancer cell survival in a stressful microenvironment, and inhibits MIA PaCa-2 spheroid formation. <i>In vivo</i>, toyaburgine, alone or combined with gemcitabine, shows effective tumor suppression in subcutaneous xenograft and clinically relevant orthotopic models, where it also reduces cachexia. These results highlight the potential of toyaburgine as a new therapeutic drug for pancreatic cancer. Its combination with gemcitabine presents a promising treatment approach by targeting both proliferating and gemcitabine-resistant cancer cells.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":" ","pages":""},"PeriodicalIF":3.5,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143565576","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":"Toyaburgine, a Synthetic N-Biphenyl-Dihydroisoquinoline Inspired by Related N,C-Coupled Naphthylisoquinoline Alkaloids, with High In Vivo Efficacy in Preclinical Pancreatic Cancer Models","authors":"Suresh Awale*,&nbsp;Juthamart Maneenet,&nbsp;Nguyen Duy Phan,&nbsp;Hung Hong Nguyen,&nbsp;Tsutomu Fujii,&nbsp;Heiko Ihmels,&nbsp;Denisa Soost,&nbsp;Nasir Tajuddeen,&nbsp;Doris Feineis and Gerhard Bringmann*,&nbsp;","doi":"10.1021/acschembio.4c0087010.1021/acschembio.4c00870","DOIUrl":"https://doi.org/10.1021/acschembio.4c00870https://doi.org/10.1021/acschembio.4c00870","url":null,"abstract":"<p >Pancreatic cancer is a highly aggressive and lethal malignancy, with a 5-year survival rate below 10%. Traditional chemotherapy, including gemcitabine, has limited efficacy due to chemoresistance and a unique tumor microenvironment characterized by hypovascularity and nutrient deprivation. This study reports on the discovery of a new <i>N</i>-biphenyl-dihydroisoquinoline, named toyaburgine (<b>4</b>), inspired by naturally occurring <i>N,C</i>-coupled naphthylisoquinoline alkaloids. Developed through systematic structural optimization, toyaburgine is a potent anticancer agent, showing promise for pancreatic cancer treatment. It exhibits strong antiausterity activity with low nanomolar PC₅₀ values, effectively inhibiting pancreatic cancer cell viability under nutrient-deprived conditions. <i>In vitro</i>, <b>4</b> causes significant morphological changes and cancer cell death in MIA PaCa-2 cells while also inhibiting cell migration and colony formation, which indicates its antimetastatic potential. Mechanistically, toyaburgine disrupts the PI3K/Akt/mTOR pathway, essential for pancreatic cancer cell survival in a stressful microenvironment, and inhibits MIA PaCa-2 spheroid formation. <i>In vivo</i>, toyaburgine, alone or combined with gemcitabine, shows effective tumor suppression in subcutaneous xenograft and clinically relevant orthotopic models, where it also reduces cachexia. These results highlight the potential of toyaburgine as a new therapeutic drug for pancreatic cancer. Its combination with gemcitabine presents a promising treatment approach by targeting both proliferating and gemcitabine-resistant cancer cells.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 4","pages":"917–929 917–929"},"PeriodicalIF":3.5,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143842323","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":"Characterization of RufT Thioesterase Domain Reveals Insights into Rufomycin Cyclization and the Biosynthetic Origin of Rufomyazine","authors":"Yaoyu Ding,&nbsp;Gustavo Perez-Ortiz,&nbsp;Alexandra-Georgiana Butulan,&nbsp;Hamzah Sharif and Sarah M. Barry*,&nbsp;","doi":"10.1021/acschembio.4c0080210.1021/acschembio.4c00802","DOIUrl":"https://doi.org/10.1021/acschembio.4c00802https://doi.org/10.1021/acschembio.4c00802","url":null,"abstract":"<p >The nonribosomal cyclic peptides (NRcPs) rufomycins, produced by <i>Streptomyces atratus</i>, have attracted attention as antimycobacterials. Thus, there has been interest in engineering the corresponding biosynthetic pathway to produce novel derivatives. We have thus investigated the type I thioesterase (TE) of the NRPS RufT that catalyzes rufomycin peptide macrocyclization to understand its tolerance to changes in substrate peptide sequence. In contrast to our previously reported efficient cyclization chemistry, the recombinant RufT-TE domain and RufT-PCP-TE didomain, while tolerating some substrate structural changes, both produce high levels of hydrolyzed peptide. Closer analysis led to the identification of the natural product diketopiperazine rufomyazine in assays. The data indicate, with significant implications for rufomycin production, that RufT produces both cyclic and linear peptides. We propose that rufomyazine forms non-enzymatically from the linear peptide. In addition, it provides evidence for TE domains as gatekeepers in NRPS biosynthesis.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 3","pages":"573–580 573–580"},"PeriodicalIF":3.5,"publicationDate":"2025-03-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acschembio.4c00802","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143667123","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":"Structure of the Outer Membrane Transporter FemA and Its Role in the Uptake of Ferric Dihydro-Aeruginoic Acid and Ferric Aeruginoic Acid in Pseudomonas aeruginosa","authors":"Virginie Will,&nbsp;Lucile Moynié*,&nbsp;Elise Si Ahmed Charrier,&nbsp;Audrey Le Bas,&nbsp;Lauriane Kuhn,&nbsp;Florian Volck,&nbsp;Johana Chicher,&nbsp;Hava Aksoy,&nbsp;Morgan Madec,&nbsp;Cyril Antheaume,&nbsp;Gaëtan L. A. Mislin and Isabelle J. Schalk*,&nbsp;","doi":"10.1021/acschembio.4c0082010.1021/acschembio.4c00820","DOIUrl":"https://doi.org/10.1021/acschembio.4c00820https://doi.org/10.1021/acschembio.4c00820","url":null,"abstract":"<p >Iron is essential for bacterial growth, and <i>Pseudomonas aeruginosa</i> synthesizes the siderophores pyochelin (PCH) and pyoverdine to acquire it. PCH contains a thiazolidine ring that aids in iron chelation but is prone to hydrolysis, leading to the formation of 2-(2-hydroxylphenyl)-thiazole-4-carbaldehyde (IQS). Using mass spectrometry, we demonstrated that PCH undergoes hydrolysis and oxidation in solution, resulting in the formation of aeruginoic acid (AA). This study used proteomic analyses and fluorescent reporters to show that AA, dihydroaeruginoic acid (DHA), and PCH induce the expression of <i>femA</i>, a gene encoding the ferri-mycobactin outer membrane transporter in <i>P. aeruginosa</i>. Notably, the induction by AA and DHA was observed only in strains unable to produce pyoverdine, suggesting their weaker iron-chelating ability compared to that of pyoverdine. ⁵⁵Fe uptake assays demonstrated that both AA-Fe and DHA-Fe complexes are transported via FemA; however, no uptake was observed for PCH-Fe through this transporter. Structural studies revealed that FemA is able to bind AA₂-Fe or DHA₂-Fe complexes. Key interactions are conserved between FemA and these two complexes, with specificity primarily driven by one of the two siderophore molecules. Interestingly, although no iron uptake was noted for PCH through FemA, the transporter also binds PCH-Fe in a similar manner. These findings show that under moderate iron deficiency, when only PCH is produced by <i>P. aeruginosa</i>, degradation products AA and DHA enhance iron uptake by inducing <i>femA</i> expression and facilitating iron transport through FemA. This provides new insights into the pathogen’s strategies for iron homeostasis.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"20 3","pages":"690–706 690–706"},"PeriodicalIF":3.5,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143667052","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}