Pub Date : 2025-02-25DOI: 10.1016/j.chembiol.2025.01.009
Tian Qiu, Saara-Anne Azizi, Shubhashree Pani, Bryan C. Dickinson
Peroxiredoxins (PRDXs) are a highly conserved family of peroxidases that serve as the primary scavengers of peroxides. Post-translational modifications play crucial roles modulating PRDX activities, tuning the balance between reactive oxygen species (ROS) signaling and stress. We previously reported that S-acylation occurs at the “peroxidatic” cysteine (Cp) site of PRDX5 and that it inhibits PRDX5 activity. Here, we show that the PRDX family more broadly is subject to S-acylation at the Cp site of all PRDXs and that PRDX S-acylation dynamically responds to cellular ROS levels. Using activity-based fluorescent imaging with DPP-Red, a red-shifted fluorescent indicator for acyl-protein thioesterase (APT) activity, we also discover that the instigation of ROS-stress via exogenous H2O2 activates both the cytosolic and mitochondrial APTs, whereas epidermal growth factor (EGF)-stimulated endogenous H2O2 deactivates the cytosolic APTs. These results indicate that APTs help tune H2O2 signal transduction and ROS protection through PRDX S-deacylation.
{"title":"Dynamic PRDX S-acylation modulates ROS stress and signaling","authors":"Tian Qiu, Saara-Anne Azizi, Shubhashree Pani, Bryan C. Dickinson","doi":"10.1016/j.chembiol.2025.01.009","DOIUrl":"https://doi.org/10.1016/j.chembiol.2025.01.009","url":null,"abstract":"Peroxiredoxins (PRDXs) are a highly conserved family of peroxidases that serve as the primary scavengers of peroxides. Post-translational modifications play crucial roles modulating PRDX activities, tuning the balance between reactive oxygen species (ROS) signaling and stress. We previously reported that <em>S</em>-acylation occurs at the “peroxidatic” cysteine (Cp) site of PRDX5 and that it inhibits PRDX5 activity. Here, we show that the PRDX family more broadly is subject to <em>S</em>-acylation at the Cp site of all PRDXs and that PRDX <em>S</em>-acylation dynamically responds to cellular ROS levels. Using activity-based fluorescent imaging with DPP-Red, a red-shifted fluorescent indicator for acyl-protein thioesterase (APT) activity, we also discover that the instigation of ROS-stress via exogenous H<sub>2</sub>O<sub>2</sub> activates both the cytosolic and mitochondrial APTs, whereas epidermal growth factor (EGF)-stimulated endogenous H<sub>2</sub>O<sub>2</sub> deactivates the cytosolic APTs. These results indicate that APTs help tune H<sub>2</sub>O<sub>2</sub> signal transduction and ROS protection through PRDX <em>S</em>-deacylation.","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"12 1","pages":""},"PeriodicalIF":8.6,"publicationDate":"2025-02-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143486527","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-24DOI: 10.1016/j.chembiol.2025.01.008
Sara Chiarugi, Francesco Margheriti, Valentina De Lorenzi, Elisa Martino, Eleonora Germana Margheritis, Aldo Moscardini, Roberto Marotta, Antonio Chaves-Sanjuan, Cristina Del Seppia, Giuseppe Federighi, Dominga Lapi, Tiziano Bandiera, Simona Rapposelli, Rossana Scuri, Martino Bolognesi, Gianpiero Garau
Thiazide and thiazide-like diuretics are among the most efficacious and used drugs for the treatment of hypertension, edema, and major cardiovascular outcomes. Despite more then than six decades of clinical use, the molecular target and mechanism of action by which these drugs cure hypertension after long-term use have remained mysterious. Here we report the discovery and validation of a previously unknown renal and extrarenal target of these antihypertensives, the membrane-associated phospholipase N-acylphosphatidylethanolamine-specific phospholipase D (NAPE-PLD) of the endocannabinoid system. Structural and functional insights, together with preclinical studies in hypertensive rats, disclose the molecular and physiological basis by which thiazides cause acute diuresis and, at the same time, the distinctive chronic reduction of vascular resistance. Our results shed light on the mechanism of treatment of hypertension and will be useful for developing more efficacious medications for the management of vascular risk factors, as well as associated leukoencephalopathies and myelin disorders.
{"title":"NAPE-PLD is target of thiazide diuretics","authors":"Sara Chiarugi, Francesco Margheriti, Valentina De Lorenzi, Elisa Martino, Eleonora Germana Margheritis, Aldo Moscardini, Roberto Marotta, Antonio Chaves-Sanjuan, Cristina Del Seppia, Giuseppe Federighi, Dominga Lapi, Tiziano Bandiera, Simona Rapposelli, Rossana Scuri, Martino Bolognesi, Gianpiero Garau","doi":"10.1016/j.chembiol.2025.01.008","DOIUrl":"https://doi.org/10.1016/j.chembiol.2025.01.008","url":null,"abstract":"Thiazide and thiazide-like diuretics are among the most efficacious and used drugs for the treatment of hypertension, edema, and major cardiovascular outcomes. Despite more then than six decades of clinical use, the molecular target and mechanism of action by which these drugs cure hypertension after long-term use have remained mysterious. Here we report the discovery and validation of a previously unknown renal and extrarenal target of these antihypertensives, the membrane-associated phospholipase N-acylphosphatidylethanolamine-specific phospholipase D (NAPE-PLD) of the endocannabinoid system. Structural and functional insights, together with preclinical studies in hypertensive rats, disclose the molecular and physiological basis by which thiazides cause acute diuresis and, at the same time, the distinctive chronic reduction of vascular resistance. Our results shed light on the mechanism of treatment of hypertension and will be useful for developing more efficacious medications for the management of vascular risk factors, as well as associated leukoencephalopathies and myelin disorders.","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"38 1","pages":""},"PeriodicalIF":8.6,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143477580","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.chembiol.2024.12.006
Jian Tang , Ramkumar Moorthy , Laura E. Hirsch , Özlem Demir , Zachary D. Baker , Jordan A. Naumann , Katherine F.M. Jones , Michael J. Grillo , Ella S. Haefner , Ke Shi , Michaella J. Levy , Harshita B. Gupta , Hideki Aihara , Reuben S. Harris , Rommie E. Amaro , Nicholas M. Levinson , Daniel A. Harki
The N-Myc transcription factor, encoded by MYCN, is a mechanistically validated, yet challenging, target for neuroblastoma (NB) therapy development. In normal neuronal progenitors, N-Myc undergoes rapid degradation, while, in MYCN-amplified NB cells, Aurora kinase A (Aurora-A) binds to and stabilizes N-Myc, resulting in elevated protein levels. Here, we demonstrate that targeted protein degradation of Aurora-A decreases N-Myc levels. A potent Aurora-A degrader, HLB-0532259 (compound 4), was developed from an Aurora-A-binding ligand that engages the Aurora-A/N-Myc complex. HLB-0532259 promotes the degradation of Aurora-A, which elicits concomitant N-Myc degradation, with nanomolar potency and excellent selectivity. HLB-0532259 surpasses the cellular efficacy of established allosteric Aurora-A inhibitors, exhibits favorable pharmacokinetic properties, and elicits tumor reduction in a murine xenograft NB model. This study broadly delineates a strategy for targeting “undruggable” proteins that are reliant on accessory proteins for cellular stabilization.
{"title":"Targeting N-Myc in neuroblastoma with selective Aurora kinase A degraders","authors":"Jian Tang , Ramkumar Moorthy , Laura E. Hirsch , Özlem Demir , Zachary D. Baker , Jordan A. Naumann , Katherine F.M. Jones , Michael J. Grillo , Ella S. Haefner , Ke Shi , Michaella J. Levy , Harshita B. Gupta , Hideki Aihara , Reuben S. Harris , Rommie E. Amaro , Nicholas M. Levinson , Daniel A. Harki","doi":"10.1016/j.chembiol.2024.12.006","DOIUrl":"10.1016/j.chembiol.2024.12.006","url":null,"abstract":"<div><div>The N-Myc transcription factor, encoded by <em>MYCN</em>, is a mechanistically validated, yet challenging, target for neuroblastoma (NB) therapy development. In normal neuronal progenitors, N-Myc undergoes rapid degradation, while, in <em>MYCN</em>-amplified NB cells, Aurora kinase A (Aurora-A) binds to and stabilizes N-Myc, resulting in elevated protein levels. Here, we demonstrate that targeted protein degradation of Aurora-A decreases N-Myc levels. A potent Aurora-A degrader, HLB-0532259 (compound <strong>4</strong>), was developed from an Aurora-A-binding ligand that engages the Aurora-A/N-Myc complex. HLB-0532259 promotes the degradation of Aurora-A, which elicits concomitant N-Myc degradation, with nanomolar potency and excellent selectivity. HLB-0532259 surpasses the cellular efficacy of established allosteric Aurora-A inhibitors, exhibits favorable pharmacokinetic properties, and elicits tumor reduction in a murine xenograft NB model. This study broadly delineates a strategy for targeting “undruggable” proteins that are reliant on accessory proteins for cellular stabilization.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 2","pages":"Pages 352-362.e10"},"PeriodicalIF":6.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142935250","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.chembiol.2024.10.001
Viviane S. De Paula , Abhinav Dubey , Haribabu Arthanari , Nikolaos G. Sgourakis
CRISPR-Cas9 has revolutionized genome engineering applications by programming its single-guide RNA, where high specificity is required. However, the precise molecular mechanism underscoring discrimination between on/off-target DNA sequences, relative to the guide RNA template, remains elusive. Here, using methyl-based NMR to study multiple holoenzymes assembled in vitro, we elucidate a discrete protein conformational state which enables recognition of DNA mismatches at the protospacer adjacent motif (PAM)-distal end. Our results delineate an allosteric pathway connecting a dynamic conformational switch at the REC3 domain, with the sampling of a catalytically competent state by the HNH domain. Our NMR data show that HiFi Cas9 (R691A) increases the fidelity of DNA recognition by stabilizing this "surveillance state" for mismatched substrates, shifting the Cas9 conformational equilibrium away from the active state. These results establish a paradigm of substrate recognition through an allosteric protein-based switch, providing unique insights into the molecular mechanism which governs Cas9 selectivity.
CRISPR-Cas9 通过对需要高特异性的单导 RNA 进行编程，彻底改变了基因组工程应用。然而，相对于引导 RNA 模板而言，区分目标 DNA 序列的精确分子机制仍未确定。在这里，我们利用基于甲基的核磁共振技术研究了体外组装的多个全酶，阐明了一种离散的蛋白质构象状态，它能识别原间隔邻接基序（PAM）远端的 DNA 错配。我们的研究结果勾勒出了一条异构途径，它将 REC3 结构域的动态构象转换与 HNH 结构域的催化状态取样连接起来。我们的核磁共振数据显示，HiFi Cas9 (R691A)通过稳定这种针对不匹配底物的 "监视状态"，使 Cas9 的构象平衡偏离活性状态，从而提高了 DNA 识别的保真度。这些结果建立了一种通过基于异构蛋白的开关来识别底物的范例，为研究支配 Cas9 选择性的分子机制提供了独特的见解。
{"title":"Dynamic sampling of a surveillance state enables DNA proofreading by Cas9","authors":"Viviane S. De Paula , Abhinav Dubey , Haribabu Arthanari , Nikolaos G. Sgourakis","doi":"10.1016/j.chembiol.2024.10.001","DOIUrl":"10.1016/j.chembiol.2024.10.001","url":null,"abstract":"<div><div>CRISPR-Cas9 has revolutionized genome engineering applications by programming its single-guide RNA, where high specificity is required. However, the precise molecular mechanism underscoring discrimination between on/off-target DNA sequences, relative to the guide RNA template, remains elusive. Here, using methyl-based NMR to study multiple holoenzymes assembled <em>in vitro</em>, we elucidate a discrete protein conformational state which enables recognition of DNA mismatches at the protospacer adjacent motif (PAM)-distal end. Our results delineate an allosteric pathway connecting a dynamic conformational switch at the REC3 domain, with the sampling of a catalytically competent state by the HNH domain. Our NMR data show that HiFi Cas9 (R691A) increases the fidelity of DNA recognition by stabilizing this \"surveillance state\" for mismatched substrates, shifting the Cas9 conformational equilibrium away from the active state. These results establish a paradigm of substrate recognition through an allosteric protein-based switch, providing unique insights into the molecular mechanism which governs Cas9 selectivity.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 2","pages":"Pages 267-279.e5"},"PeriodicalIF":6.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142519849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.chembiol.2024.12.001
Bradley E. Poulsen , Thulasi Warrier , Sulyman Barkho , Josephine Bagnall , Keith P. Romano , Tiantian White , Xiao Yu , Tomohiko Kawate , Phuong H. Nguyen , Kyra Raines , Kristina Ferrara , A. Lorelei Golas , Michael FitzGerald , Andras Boeszoermenyi , Virendar Kaushik , Michael Serrano-Wu , Noam Shoresh , Deborah T. Hung
The surge of antimicrobial resistance threatens efficacy of current antibiotics, particularly against Pseudomonas aeruginosa, a highly resistant gram-negative pathogen. The asymmetric outer membrane (OM) of P. aeruginosa combined with its array of efflux pumps provide a barrier to xenobiotic accumulation, thus making antibiotic discovery challenging. We adapted PROSPECT, a target-based, whole-cell screening strategy, to discover small molecule probes that kill P. aeruginosa mutants depleted for essential proteins localized at the OM. We identified BRD1401, a small molecule that has specific activity against a P. aeruginosa mutant depleted for the essential lipoprotein, OprL. Genetic and chemical biological studies identified that BRD1401 acts by targeting the OM β-barrel protein OprH to disrupt its interaction with LPS and increase membrane fluidity. Studies with BRD1401 also revealed an interaction between OprL and OprH, directly linking the OM with peptidoglycan. Thus, a whole-cell, multiplexed screen can identify species-specific chemical probes to reveal pathogen biology.
{"title":"Discovery of a Pseudomonas aeruginosa-specific small molecule targeting outer membrane protein OprH-LPS interaction by a multiplexed screen","authors":"Bradley E. Poulsen , Thulasi Warrier , Sulyman Barkho , Josephine Bagnall , Keith P. Romano , Tiantian White , Xiao Yu , Tomohiko Kawate , Phuong H. Nguyen , Kyra Raines , Kristina Ferrara , A. Lorelei Golas , Michael FitzGerald , Andras Boeszoermenyi , Virendar Kaushik , Michael Serrano-Wu , Noam Shoresh , Deborah T. Hung","doi":"10.1016/j.chembiol.2024.12.001","DOIUrl":"10.1016/j.chembiol.2024.12.001","url":null,"abstract":"<div><div>The surge of antimicrobial resistance threatens efficacy of current antibiotics, particularly against <em>Pseudomonas aeruginosa</em>, a highly resistant gram-negative pathogen. The asymmetric outer membrane (OM) of <em>P. aeruginosa</em> combined with its array of efflux pumps provide a barrier to xenobiotic accumulation, thus making antibiotic discovery challenging. We adapted PROSPECT, a target-based, whole-cell screening strategy, to discover small molecule probes that kill <em>P. aeruginosa</em> mutants depleted for essential proteins localized at the OM. We identified BRD1401, a small molecule that has specific activity against a <em>P. aeruginosa</em> mutant depleted for the essential lipoprotein, OprL. Genetic and chemical biological studies identified that BRD1401 acts by targeting the OM β-barrel protein OprH to disrupt its interaction with LPS and increase membrane fluidity. Studies with BRD1401 also revealed an interaction between OprL and OprH, directly linking the OM with peptidoglycan. Thus, a whole-cell, multiplexed screen can identify species-specific chemical probes to reveal pathogen biology.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 2","pages":"Pages 307-324.e15"},"PeriodicalIF":6.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142888682","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.chembiol.2024.12.013
Cong Shen , Aldo I. Salazar-Morales , Wonhyeuk Jung , Joey Erwin , Yangqi Gu , Anthony Coelho , Kallol Gupta , Sibel Ebru Yalcin , Fadel A. Samatey , Nikhil S. Malvankar
Microbial extracellular electron transfer (EET) drives various globally important environmental phenomena and has biotechnology applications. Diverse prokaryotes have been proposed to perform EET via surface-displayed “nanowires” composed of multi-heme cytochromes. However, the mechanism that enables only a few cytochromes to polymerize into nanowires is unclear. Here, we identify a highly conserved omcS-companion (osc) cluster that drives the formation of cytochrome OmcS nanowires in Geobacter sulfurreducens. Through a combination of genetic, biochemical, and biophysical methods, we establish that prolyl isomerase-containing chaperon OscH, channel-like OscEFG, and β-propeller-like OscD are involved in the folding, secretion, and morphology maintenance of OmcS nanowires, respectively. OscH and OscG can interact with OmcS. Furthermore, overexpression of oscG accelerates EET by overproducing nanowires in an ATP-dependent manner. Heme loading splits OscD; ΔoscD accelerates cell growth, bundles nanowires into cables. Our findings establish the mechanism and prevalence of a specialized and modular assembly system for nanowires across phylogenetically diverse species and environments
{"title":"A widespread and ancient bacterial machinery assembles cytochrome OmcS nanowires essential for extracellular electron transfer","authors":"Cong Shen , Aldo I. Salazar-Morales , Wonhyeuk Jung , Joey Erwin , Yangqi Gu , Anthony Coelho , Kallol Gupta , Sibel Ebru Yalcin , Fadel A. Samatey , Nikhil S. Malvankar","doi":"10.1016/j.chembiol.2024.12.013","DOIUrl":"10.1016/j.chembiol.2024.12.013","url":null,"abstract":"<div><div>Microbial extracellular electron transfer (EET) drives various globally important environmental phenomena and has biotechnology applications. Diverse prokaryotes have been proposed to perform EET via surface-displayed “nanowires” composed of multi-heme cytochromes. However, the mechanism that enables only a few cytochromes to polymerize into nanowires is unclear. Here, we identify a highly conserved <em><u>o</u>mc<u>S</u></em>-<u>c</u>ompanion (<em>osc</em>) cluster that drives the formation of cytochrome OmcS nanowires in <em>Geobacter sulfurreducens</em>. Through a combination of genetic, biochemical, and biophysical methods, we establish that prolyl isomerase-containing chaperon OscH, channel-like OscEFG, and β-propeller-like OscD are involved in the folding, secretion, and morphology maintenance of OmcS nanowires, respectively. OscH and OscG can interact with OmcS. Furthermore, overexpression of <em>oscG</em> accelerates EET by overproducing nanowires in an ATP-dependent manner. Heme loading splits OscD; Δ<em>oscD</em> accelerates cell growth, bundles nanowires into cables. Our findings establish the mechanism and prevalence of a specialized and modular assembly system for nanowires across phylogenetically diverse species and environments</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 2","pages":"Pages 239-254.e7"},"PeriodicalIF":6.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142981366","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.chembiol.2025.01.002
Joseph J. Smith , Taylor R. Valentino , Austin H. Ablicki , Riddhidev Banerjee , Adam R. Colligan , Debra M. Eckert , Gabrielle A. Desjardins , Katharine L. Diehl
Acetyl-coenzyme A is a central metabolite that participates in many cellular pathways. Evidence suggests that acetyl-CoA metabolism is highly compartmentalized in mammalian cells. Yet methods to measure acetyl-CoA in living cells are lacking. Herein, we engineered an acetyl-CoA biosensor from the bacterial protein PanZ and circularly permuted green fluorescent protein (cpGFP). The sensor, “PancACe,” has a maximum change of ∼2-fold and a response range of ∼10 μM–2 mM acetyl-CoA. We demonstrated that the sensor has a greater than 7-fold selectivity over coenzyme A, butyryl-CoA, malonyl-CoA, and succinyl-CoA, and a 2.3-fold selectivity over propionyl-CoA. We expressed the sensor in E. coli and showed that it enables detection of rapid changes in acetyl-CoA levels. By localizing the sensor to either the cytoplasm, nucleus, or mitochondria in human cells, we showed that it enables subcellular detection of changes in acetyl-CoA levels, the magnitudes of which agreed with an orthogonal PicoProbe assay.
{"title":"A genetically encoded fluorescent biosensor for visualization of acetyl-CoA in live cells","authors":"Joseph J. Smith , Taylor R. Valentino , Austin H. Ablicki , Riddhidev Banerjee , Adam R. Colligan , Debra M. Eckert , Gabrielle A. Desjardins , Katharine L. Diehl","doi":"10.1016/j.chembiol.2025.01.002","DOIUrl":"10.1016/j.chembiol.2025.01.002","url":null,"abstract":"<div><div>Acetyl-coenzyme A is a central metabolite that participates in many cellular pathways. Evidence suggests that acetyl-CoA metabolism is highly compartmentalized in mammalian cells. Yet methods to measure acetyl-CoA in living cells are lacking. Herein, we engineered an acetyl-CoA biosensor from the bacterial protein PanZ and circularly permuted green fluorescent protein (cpGFP). The sensor, “PancACe,” has a maximum change of ∼2-fold and a response range of ∼10 μM–2 mM acetyl-CoA. We demonstrated that the sensor has a greater than 7-fold selectivity over coenzyme A, butyryl-CoA, malonyl-CoA, and succinyl-CoA, and a 2.3-fold selectivity over propionyl-CoA. We expressed the sensor in <em>E. coli</em> and showed that it enables detection of rapid changes in acetyl-CoA levels. By localizing the sensor to either the cytoplasm, nucleus, or mitochondria in human cells, we showed that it enables subcellular detection of changes in acetyl-CoA levels, the magnitudes of which agreed with an orthogonal PicoProbe assay.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 2","pages":"Pages 325-337.e10"},"PeriodicalIF":6.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143044669","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.chembiol.2025.01.004
Hong-Yi Zhao , Zhongwei Liu , Jinsong Tao , Shuai Mao , Meilin Wang , Miao He , Bo Wen , Wei Gao , Duxin Sun
Stimulator of interferon genes (STING) agonists could overcome immunosuppressive microenvironment to improve cancer immunotherapy. However, it is challenging to develop oral STING agonists to achieve systemic immunity. In this study, we discovered ZSA-51 as a potent oral STING agonist with distinct benzo[4,5]thieno[2,3-c]pyrrole-1,3-dione scaffold through tricyclic scaffold screening. ZSA-51, as a prodrug, exhibited nanomolar in vitro STING activation activity and potent in vivo antitumor efficacy in both colon and pancreatic cancer models. The specificity of ZSA-51 in activating STING was confirmed using STING knockout cells and a structurally similar but negative control compound. Moreover, ZSA-51 demonstrated superior oral pharmacokinetic (PK) properties with low toxicity. Importantly, ZSA-51 remodeled immune microenvironment both in tumor and lymph node. Our data suggest that ZSA-51 is a potent oral STING agonist with robust anticancer efficacy, superior PK properties, and low toxicity, holding potential for future development for cancer immunotherapy.
{"title":"An oral tricyclic STING agonist suppresses tumor growth through remodeling of the immune microenvironment","authors":"Hong-Yi Zhao , Zhongwei Liu , Jinsong Tao , Shuai Mao , Meilin Wang , Miao He , Bo Wen , Wei Gao , Duxin Sun","doi":"10.1016/j.chembiol.2025.01.004","DOIUrl":"10.1016/j.chembiol.2025.01.004","url":null,"abstract":"<div><div>Stimulator of interferon genes (STING) agonists could overcome immunosuppressive microenvironment to improve cancer immunotherapy. However, it is challenging to develop oral STING agonists to achieve systemic immunity. In this study, we discovered ZSA-51 as a potent oral STING agonist with distinct benzo[4,5]thieno[2,3-<em>c</em>]pyrrole-1,3-dione scaffold through tricyclic scaffold screening. ZSA-51, as a prodrug, exhibited nanomolar <em>in vitro</em> STING activation activity and potent <em>in vivo</em> antitumor efficacy in both colon and pancreatic cancer models. The specificity of ZSA-51 in activating STING was confirmed using STING knockout cells and a structurally similar but negative control compound. Moreover, ZSA-51 demonstrated superior oral pharmacokinetic (PK) properties with low toxicity. Importantly, ZSA-51 remodeled immune microenvironment both in tumor and lymph node. Our data suggest that ZSA-51 is a potent oral STING agonist with robust anticancer efficacy, superior PK properties, and low toxicity, holding potential for future development for cancer immunotherapy.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 2","pages":"Pages 280-290.e14"},"PeriodicalIF":6.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143077214","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.chembiol.2024.10.008
Criseyda Martinez , Yan Xiong , Alison Bartkowski , Ibuki Harada , Xiaoxiao Ren , Jessica Byerly , Elisa Port , Jian Jin , Hanna Irie
Protein tyrosine kinase 6 (PTK6), a non-receptor tyrosine kinase, is an oncogenic driver in many tumor types. However, agents that therapeutically target PTK6 are lacking. Although several PTK6 kinase inhibitors have been developed, none have been clinically translated, which may be due to kinase-independent functions that compromise their efficacy. PTK6 kinase inhibitor treatment phenocopies some, but not all effects of PTK6 downregulation. PTK6 downregulation inhibits growth of breast cancer cells, but treatment with PTK6 kinase inhibitor does not. To chemically downregulate PTK6, we designed a PROTAC, MS105, which potently and specifically degrades PTK6. Treatment with MS105, but not PTK6 kinase inhibitor, inhibits growth and induces apoptosis of breast cancer cells, phenocopying the effects of PTK6 (short hairpin RNA) shRNA/CRISPR. In contrast, both MS105 and PTK6 kinase inhibitor effectively inhibit breast cancer cell migration, supporting the differing kinase dependencies of PTK6’s oncogenic functions. Our studies support PTK6 degraders as a preferred approach to targeting PTK6 in cancer.
{"title":"A PROTAC degrader suppresses oncogenic functions of PTK6, inducing apoptosis of breast cancer cells","authors":"Criseyda Martinez , Yan Xiong , Alison Bartkowski , Ibuki Harada , Xiaoxiao Ren , Jessica Byerly , Elisa Port , Jian Jin , Hanna Irie","doi":"10.1016/j.chembiol.2024.10.008","DOIUrl":"10.1016/j.chembiol.2024.10.008","url":null,"abstract":"<div><div>Protein tyrosine kinase 6 (PTK6), a non-receptor tyrosine kinase, is an oncogenic driver in many tumor types. However, agents that therapeutically target PTK6 are lacking. Although several PTK6 kinase inhibitors have been developed, none have been clinically translated, which may be due to kinase-independent functions that compromise their efficacy. PTK6 kinase inhibitor treatment phenocopies some, but not all effects of PTK6 downregulation. PTK6 downregulation inhibits growth of breast cancer cells, but treatment with PTK6 kinase inhibitor does not. To chemically downregulate PTK6, we designed a PROTAC, MS105, which potently and specifically degrades PTK6. Treatment with MS105, but not PTK6 kinase inhibitor, inhibits growth and induces apoptosis of breast cancer cells, phenocopying the effects of PTK6 (short hairpin RNA) shRNA/CRISPR. In contrast, both MS105 and PTK6 kinase inhibitor effectively inhibit breast cancer cell migration, supporting the differing kinase dependencies of PTK6’s oncogenic functions. Our studies support PTK6 degraders as a preferred approach to targeting PTK6 in cancer.</div></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"32 2","pages":"Pages 255-266.e8"},"PeriodicalIF":6.6,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142601055","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：