Pub Date : 2024-08-15DOI: 10.1016/j.chembiol.2024.03.004
Adhesion G protein-coupled receptor (aGPCR) signaling influences development and homeostasis in a wide range of tissues. In the current model for aGPCR signaling, ligand binding liberates a conserved sequence that acts as an intramolecular, tethered agonist (TA), yet this model has not been evaluated systematically for all aGPCRs. Here, we assessed the TA-dependent activities of all 33 aGPCRs in a suite of transcriptional reporter, G protein activation, and β-arrestin recruitment assays using a new fusion protein platform. Strikingly, only ∼50% of aGPCRs exhibited robust TA-dependent activation, and unlike other GPCR families, aGPCRs showed a notable preference for G12/13 signaling. AlphaFold2 predictions assessing TA engagement in the predicted intramolecular binding pocket aligned with the TA dependence of the cellular responses. This dataset provides a comprehensive resource to inform the investigation of all human aGPCRs and for targeting aGPCRs therapeutically.
粘附 G 蛋白偶联受体(aGPCR)信号传导影响着多种组织的发育和稳态。在目前的 aGPCR 信号传导模型中,配体结合会释放出一个保守序列,该序列可充当分子内的系链激动剂(TA),但这一模型尚未针对所有 aGPCR 进行过系统评估。在这里,我们利用一个新的融合蛋白平台,在一系列转录报告、G 蛋白激活和 β - 逮捕素招募试验中评估了所有 33 个 aGPCR 的 TA 依赖性活性。令人吃惊的是,只有 50% 的 aGPCR 表现出强大的 TA 依赖性激活,而且与其他 GPCR 家族不同,aGPCR 对 G12/13 信号转导表现出明显的偏好。AlphaFold2 预测评估了 TA 在预测的分子内结合口袋中的参与情况,这与细胞反应的 TA 依赖性相一致。该数据集提供了一个全面的资源,为研究所有人类 aGPCRs 和治疗 aGPCRs 提供了信息。
{"title":"Heterogeneity of tethered agonist signaling in adhesion G protein-coupled receptors","authors":"","doi":"10.1016/j.chembiol.2024.03.004","DOIUrl":"10.1016/j.chembiol.2024.03.004","url":null,"abstract":"<div><p><span><span><span>Adhesion G protein-coupled receptor (aGPCR) signaling influences development and homeostasis<span> in a wide range of tissues. In the current model for aGPCR signaling, ligand binding liberates a </span></span>conserved sequence<span> that acts as an intramolecular, tethered agonist (TA), yet this model has not been evaluated systematically for all aGPCRs. Here, we assessed the TA-dependent activities of all 33 aGPCRs in a suite of transcriptional reporter, G protein activation, and β-arrestin recruitment assays using a new </span></span>fusion protein platform. Strikingly, only ∼50% of aGPCRs exhibited robust TA-dependent activation, and unlike other GPCR families, aGPCRs showed a notable preference for G</span><sub>12/13</sub> signaling. AlphaFold2 predictions assessing TA engagement in the predicted intramolecular binding pocket aligned with the TA dependence of the cellular responses. This dataset provides a comprehensive resource to inform the investigation of all human aGPCRs and for targeting aGPCRs therapeutically.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1542-1553.e4"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140622936","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 : 2024-08-15DOI: 10.1016/j.chembiol.2024.07.013
Kyle Mangano , Patrick Ryan Potts
In this issue of Cell Chemical Biology, Raina et al.1 demonstrate proof of concept of a new chemical induced proximity strategy for targeted cancer therapeutics. Building on a recent surge in induced proximity modalities, RIPTACs represent a novel approach that offers promise in treating cancers with improved safety profiles.
在本期《细胞化学生物学》(Cell Chemical Biology)杂志上,Raina 等人1 展示了一种新型化学诱导接近策略的概念验证,该策略可用于癌症靶向治疗。RIPTACs 是一种新型的诱导接近模式,它以最近激增的诱导接近模式为基础,有望在治疗癌症的同时提高安全性。
{"title":"Feel the breeze: Opening the therapeutic window with RIPTACs and induced proximity","authors":"Kyle Mangano , Patrick Ryan Potts","doi":"10.1016/j.chembiol.2024.07.013","DOIUrl":"10.1016/j.chembiol.2024.07.013","url":null,"abstract":"<div><p>In this issue of <em>Cell Chemical Biology</em>, Raina et al.<span><span><sup>1</sup></span></span> demonstrate proof of concept of a new chemical induced proximity strategy for targeted cancer therapeutics. Building on a recent surge in induced proximity modalities, RIPTACs represent a novel approach that offers promise in treating cancers with improved safety profiles.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1391-1393"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141990566","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 : 2024-08-15DOI: 10.1016/j.chembiol.2024.07.001
Aloysus Lawong , Suraksha Gahalawat , Sneha Ray , Nhi Ho , Yan Han , Kurt E. Ward , Xiaoyi Deng , Zhe Chen , Ashwani Kumar , Chao Xing , Varun Hosangadi , Kate J. Fairhurst , Kyuto Tashiro , Glen Liszczak , David M. Shackleford , Kasiram Katneni , Gong Chen , Jessica Saunders , Elly Crighton , Arturo Casas , Margaret A. Phillips
Malaria remains a global health concern as drug resistance threatens treatment programs. We identified a piperidine carboxamide (SW042) with anti-malarial activity by phenotypic screening. Selection of SW042-resistant Plasmodium falciparum (Pf) parasites revealed point mutations in the Pf_proteasome β5 active-site (Pfβ5). A potent analog (SW584) showed efficacy in a mouse model of human malaria after oral dosing. SW584 had a low propensity to generate resistance (minimum inoculum for resistance [MIR] >109) and was synergistic with dihydroartemisinin. Pf_proteasome purification was facilitated by His8-tag introduction onto β7. Inhibition of Pfβ5 correlated with parasite killing, without inhibiting human proteasome isoforms or showing cytotoxicity. The Pf_proteasome_SW584 cryoelectron microscopy (cryo-EM) structure showed that SW584 bound non-covalently distal from the catalytic threonine, in an unexplored pocket at the β5/β6/β3 subunit interface that has species differences between Pf and human proteasomes. Identification of a reversible, species selective, orally active series with low resistance propensity provides a path for drugging this essential target.
{"title":"Identification of potent and reversible piperidine carboxamides that are species-selective orally active proteasome inhibitors to treat malaria","authors":"Aloysus Lawong , Suraksha Gahalawat , Sneha Ray , Nhi Ho , Yan Han , Kurt E. Ward , Xiaoyi Deng , Zhe Chen , Ashwani Kumar , Chao Xing , Varun Hosangadi , Kate J. Fairhurst , Kyuto Tashiro , Glen Liszczak , David M. Shackleford , Kasiram Katneni , Gong Chen , Jessica Saunders , Elly Crighton , Arturo Casas , Margaret A. Phillips","doi":"10.1016/j.chembiol.2024.07.001","DOIUrl":"10.1016/j.chembiol.2024.07.001","url":null,"abstract":"<div><p>Malaria remains a global health concern as drug resistance threatens treatment programs. We identified a piperidine carboxamide (SW042) with anti-malarial activity by phenotypic screening. Selection of SW042-resistant <em>Plasmodium falciparum</em> (<em>Pf</em>) parasites revealed point mutations in the <em>Pf_</em>proteasome β5 active-site (<em>Pf</em>β5). A potent analog (SW584) showed efficacy in a mouse model of human malaria after oral dosing. SW584 had a low propensity to generate resistance (minimum inoculum for resistance [MIR] >10<sup>9</sup>) and was synergistic with dihydroartemisinin. <em>Pf_</em>proteasome purification was facilitated by His<sub>8</sub>-tag introduction onto β7. Inhibition of <em>Pf</em>β5 correlated with parasite killing, without inhibiting human proteasome isoforms or showing cytotoxicity. The <em>Pf_</em>proteasome_SW584 cryoelectron microscopy (cryo-EM) structure showed that SW584 bound non-covalently distal from the catalytic threonine, in an unexplored pocket at the β5/β6/β3 subunit interface that has species differences between <em>Pf</em> and human proteasomes. Identification of a reversible, species selective, orally active series with low resistance propensity provides a path for drugging this essential target.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1503-1517.e19"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2451945624002782/pdfft?md5=d0d63c5b95768cd07bd4ad29366f279e&pid=1-s2.0-S2451945624002782-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141795066","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 : 2024-08-15DOI: 10.1016/j.chembiol.2024.07.007
Goldie V. Roth , Isabella R. Gengaro , Lei S. Qi
The epigenome is a complex framework through which gene expression is precisely and flexibly modulated to incorporate heritable memory and responses to environmental stimuli. It governs diverse cellular processes, including cell fate, disease, and aging. The need to understand this system and precisely control gene expression outputs for therapeutic purposes has precipitated the development of a diverse set of epigenetic editing tools. Here, we review the existing toolbox for targeted epigenetic editing, technical considerations of the current technologies, and opportunities for future development. We describe applications of therapeutic epigenetic editing and their potential for treating disease, with a discussion of ongoing delivery challenges that impede certain clinical interventions, particularly in the brain. With simultaneous advancements in available engineering tools and appropriate delivery technologies, we predict that epigenetic editing will increasingly cement itself as a powerful approach for safely treating a wide range of disorders in all tissues of the body.
{"title":"Precision epigenetic editing: Technological advances, enduring challenges, and therapeutic applications","authors":"Goldie V. Roth , Isabella R. Gengaro , Lei S. Qi","doi":"10.1016/j.chembiol.2024.07.007","DOIUrl":"10.1016/j.chembiol.2024.07.007","url":null,"abstract":"<div><p>The epigenome is a complex framework through which gene expression is precisely and flexibly modulated to incorporate heritable memory and responses to environmental stimuli. It governs diverse cellular processes, including cell fate, disease, and aging. The need to understand this system and precisely control gene expression outputs for therapeutic purposes has precipitated the development of a diverse set of epigenetic editing tools. Here, we review the existing toolbox for targeted epigenetic editing, technical considerations of the current technologies, and opportunities for future development. We describe applications of therapeutic epigenetic editing and their potential for treating disease, with a discussion of ongoing delivery challenges that impede certain clinical interventions, particularly in the brain. With simultaneous advancements in available engineering tools and appropriate delivery technologies, we predict that epigenetic editing will increasingly cement itself as a powerful approach for safely treating a wide range of disorders in all tissues of the body.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1422-1446"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S245194562400309X/pdfft?md5=d1b75cb9f1e6f7156ed373d1c1fdaec2&pid=1-s2.0-S245194562400309X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141918647","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 : 2024-08-15DOI: 10.1016/j.chembiol.2024.07.008
Xueqin Jin , Jian Huang , Huan Wang , Kan Wang , Nieng Yan
Voltage-gated sodium (Nav) and calcium (Cav) channels are responsible for the initiation of electrical signals. They have long been targeted for the treatment of various diseases. The mounting number of cryoelectron microscopy (cryo-EM) structures for diverse subtypes of Nav and Cav channels from multiple organisms necessitates a generic residue numbering system to establish the structure-function relationship and to aid rational drug design or optimization. Here we suggest a structure-based residue numbering scheme, centering around the most conserved residues on each of the functional segments. We elaborate the generic numbers through illustrative examples, focusing on representative drug-binding sites of eukaryotic Nav and Cav channels. We also extend the numbering scheme to compare common disease mutations among different Nav subtypes. Application of the generic residue numbering scheme affords immediate insights into hotspots for pathogenic mutations and critical loci for drug binding and will facilitate drug discovery targeting Nav and Cav channels.
电压门控钠(Nav)和钙(Cav)通道负责启动电信号。长期以来,它们一直是治疗各种疾病的靶标。来自多种生物体的不同亚型 Nav 和 Cav 通道的冷冻电子显微镜(cryo-EM)结构越来越多,这就需要一个通用的残基编号系统来建立结构-功能关系,并帮助合理的药物设计或优化。在此,我们围绕每个功能片段上最保守的残基,提出了基于结构的残基编号方案。我们以真核生物 Nav 和 Cav 通道的代表性药物结合位点为例,详细阐述了通用编号。我们还扩展了编号方案,以比较不同 Nav 亚型之间的常见疾病突变。应用通用残基编号方案可立即了解致病突变的热点和药物结合的关键位点,并将促进针对 Nav 和 Cav 通道的药物发现。
{"title":"A versatile residue numbering scheme for Nav and Cav channels","authors":"Xueqin Jin , Jian Huang , Huan Wang , Kan Wang , Nieng Yan","doi":"10.1016/j.chembiol.2024.07.008","DOIUrl":"10.1016/j.chembiol.2024.07.008","url":null,"abstract":"<div><p>Voltage-gated sodium (Na<sub>v</sub>) and calcium (Ca<sub>v</sub>) channels are responsible for the initiation of electrical signals. They have long been targeted for the treatment of various diseases. The mounting number of cryoelectron microscopy (cryo-EM) structures for diverse subtypes of Na<sub>v</sub> and Ca<sub>v</sub> channels from multiple organisms necessitates a generic residue numbering system to establish the structure-function relationship and to aid rational drug design or optimization. Here we suggest a structure-based residue numbering scheme, centering around the most conserved residues on each of the functional segments. We elaborate the generic numbers through illustrative examples, focusing on representative drug-binding sites of eukaryotic Na<sub>v</sub> and Ca<sub>v</sub> channels. We also extend the numbering scheme to compare common disease mutations among different Na<sub>v</sub> subtypes. Application of the generic residue numbering scheme affords immediate insights into hotspots for pathogenic mutations and critical loci for drug binding and will facilitate drug discovery targeting Na<sub>v</sub> and Ca<sub>v</sub> channels.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1394-1404"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141990567","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 : 2024-08-15DOI: 10.1016/j.chembiol.2024.07.016
Thomas R. Cech, Emmanuelle Charpentier, Aaron Ciechanover, Robert J. Lefkowitz, Kurt Wüthrich
Since the first award in 1901, the Nobel Prize has come to signify the pinnacle of scientific achievement. In this Voices piece in the August special issue of Cell Chemical Biology entitled “Bridging chemistry and biology,” we ask Nobel laureates to reflect on the impact the prize had on them. We learn how it affected their life or work, their outlook on science, the lessons learned, and their advice for the next generation of scientists.
{"title":"Reflections from Nobel laureates in chemistry","authors":"Thomas R. Cech, Emmanuelle Charpentier, Aaron Ciechanover, Robert J. Lefkowitz, Kurt Wüthrich","doi":"10.1016/j.chembiol.2024.07.016","DOIUrl":"10.1016/j.chembiol.2024.07.016","url":null,"abstract":"<div><p>Since the first award in 1901, the Nobel Prize has come to signify the pinnacle of scientific achievement. In this Voices piece in the August special issue of <em>Cell Chemical Biology</em> entitled “Bridging chemistry and biology,” we ask Nobel laureates to reflect on the impact the prize had on them. We learn how it affected their life or work, their outlook on science, the lessons learned, and their advice for the next generation of scientists.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1388-1390"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141990569","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 : 2024-08-15DOI: 10.1016/j.chembiol.2024.06.001
Synthetic biology aims to engineer complex biological systems using modular elements, with coiled-coil (CC) dimer-forming modules are emerging as highly useful building blocks in the regulation of protein assemblies and biological processes. Those small modules facilitate highly specific and orthogonal protein-protein interactions, offering versatility for the regulation of diverse biological functions. Additionally, their design rules enable precise control and tunability over these interactions, which are crucial for specific applications. Recent advancements showcase their potential for use in innovative therapeutic interventions and biomedical applications. In this review, we discuss the potential of CCs, exploring their diverse applications in mammalian cells, such as synthetic biological circuit design, transcriptional and allosteric regulation, cellular assemblies, chimeric antigen receptor (CAR) T cell regulation, and genome editing and their role in advancing the understanding and regulation of cellular processes.
合成生物学旨在利用模块化元素设计复杂的生物系统,而形成盘绕线圈(CC)二聚体的模块正在成为调控蛋白质组装和生物过程的非常有用的构件。这些小模块可促进蛋白质与蛋白质之间高度特异和正交的相互作用,为调控各种生物功能提供了多功能性。此外,它们的设计规则能够实现对这些相互作用的精确控制和可调谐性,这对特定应用至关重要。最近的进展展示了它们在创新治疗干预和生物医学应用方面的潜力。在这篇综述中,我们将讨论 CCs 的潜力,探讨它们在哺乳动物细胞中的各种应用,如合成生物电路设计、转录和异生调控、细胞组装、嵌合抗原受体 (CAR) T 细胞调控和基因组编辑,以及它们在促进对细胞过程的理解和调控方面的作用。
{"title":"The art of designed coiled-coils for the regulation of mammalian cells","authors":"","doi":"10.1016/j.chembiol.2024.06.001","DOIUrl":"10.1016/j.chembiol.2024.06.001","url":null,"abstract":"<div><p>Synthetic biology aims to engineer complex biological systems using modular elements, with coiled-coil (CC) dimer-forming modules are emerging as highly useful building blocks in the regulation of protein assemblies and biological processes. Those small modules facilitate highly specific and orthogonal protein-protein interactions, offering versatility for the regulation of diverse biological functions. Additionally, their design rules enable precise control and tunability over these interactions, which are crucial for specific applications. Recent advancements showcase their potential for use in innovative therapeutic interventions and biomedical applications. In this review, we discuss the potential of CCs, exploring their diverse applications in mammalian cells, such as synthetic biological circuit design, transcriptional and allosteric regulation, cellular assemblies, chimeric antigen receptor (CAR) T cell regulation, and genome editing and their role in advancing the understanding and regulation of cellular processes.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1460-1472"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2451945624002204/pdfft?md5=85fef3f748d2012ce1eea38d5efda70f&pid=1-s2.0-S2451945624002204-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141544253","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 : 2024-08-15DOI: 10.1016/j.chembiol.2024.05.002
Aspartate is crucial for nucleotide synthesis, ammonia detoxification, and maintaining redox balance via the malate-aspartate-shuttle (MAS). To disentangle these multiple roles of aspartate metabolism, tools are required that measure aspartate concentrations in real time and in live cells. We introduce AspSnFR, a genetically encoded green fluorescent biosensor for intracellular aspartate, engineered through displaying and screening biosensor libraries on mammalian cells. In live cells, AspSnFR is able to precisely and quantitatively measure cytosolic aspartate concentrations and dissect its production from glutamine. Combining high-content imaging of AspSnFR with pharmacological perturbations exposes differences in metabolic vulnerabilities of aspartate levels based on nutrient availability. Further, AspSnFR facilitates tracking of aspartate export from mitochondria through SLC25A12, the MAS’ key transporter. We show that SLC25A12 is a rapidly responding and direct route to couple Ca2+ signaling with mitochondrial aspartate export. This establishes SLC25A12 as a crucial link between cellular signaling, mitochondrial respiration, and metabolism.
{"title":"AspSnFR: A genetically encoded biosensor for real-time monitoring of aspartate in live cells","authors":"","doi":"10.1016/j.chembiol.2024.05.002","DOIUrl":"10.1016/j.chembiol.2024.05.002","url":null,"abstract":"<div><p>Aspartate is crucial for nucleotide synthesis, ammonia detoxification, and maintaining redox balance via the malate-aspartate-shuttle (MAS). To disentangle these multiple roles of aspartate metabolism, tools are required that measure aspartate concentrations in real time and in live cells. We introduce AspSnFR, a genetically encoded green fluorescent biosensor for intracellular aspartate, engineered through displaying and screening biosensor libraries on mammalian cells. In live cells, AspSnFR is able to precisely and quantitatively measure cytosolic aspartate concentrations and dissect its production from glutamine. Combining high-content imaging of AspSnFR with pharmacological perturbations exposes differences in metabolic vulnerabilities of aspartate levels based on nutrient availability. Further, AspSnFR facilitates tracking of aspartate export from mitochondria through SLC25A12, the MAS’ key transporter. We show that SLC25A12 is a rapidly responding and direct route to couple Ca<sup>2+</sup> signaling with mitochondrial aspartate export. This establishes SLC25A12 as a crucial link between cellular signaling, mitochondrial respiration, and metabolism.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1529-1541.e12"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S245194562400179X/pdfft?md5=3b3153c35af18753feadccb80f236687&pid=1-s2.0-S245194562400179X-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141156564","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 : 2024-08-15DOI: 10.1016/j.chembiol.2024.07.003
Dominik Brokatzky , Margarida C. Gomes , Stevens Robertin , Carolina Albino , Sydney L. Miles , Serge Mostowy
The septin cytoskeleton is primarily known for roles in cell division and host defense against bacterial infection. Despite recent insights, the full breadth of roles for septins in host defense is poorly understood. In macrophages, Shigella induces pyroptosis, a pro-inflammatory form of cell death dependent upon gasdermin D (GSDMD) pores at the plasma membrane and cell surface protein ninjurin-1 (NINJ1) for membrane rupture. Here, we discover that septins promote macrophage pyroptosis induced by lipopolysaccharide (LPS)/nigericin and Shigella infection, but do not affect cytokine expression or release. We observe that septin filaments assemble at the plasma membrane, and cleavage of GSDMD is impaired in septin-depleted cells. We found that septins regulate mitochondrial dynamics and the expression of NINJ1. Using a Shigella-zebrafish infection model, we show that septin-mediated pyroptosis is an in vivo mechanism of infection control. The discovery of septins as a mediator of pyroptosis may inspire innovative anti-bacterial and anti-inflammatory treatments.
{"title":"Septins promote macrophage pyroptosis by regulating gasdermin D cleavage and ninjurin-1-mediated plasma membrane rupture","authors":"Dominik Brokatzky , Margarida C. Gomes , Stevens Robertin , Carolina Albino , Sydney L. Miles , Serge Mostowy","doi":"10.1016/j.chembiol.2024.07.003","DOIUrl":"10.1016/j.chembiol.2024.07.003","url":null,"abstract":"<div><p>The septin cytoskeleton is primarily known for roles in cell division and host defense against bacterial infection. Despite recent insights, the full breadth of roles for septins in host defense is poorly understood. In macrophages, <em>Shigella</em> induces pyroptosis, a pro-inflammatory form of cell death dependent upon gasdermin D (GSDMD) pores at the plasma membrane and cell surface protein ninjurin-1 (NINJ1) for membrane rupture. Here, we discover that septins promote macrophage pyroptosis induced by lipopolysaccharide (LPS)/nigericin and <em>Shigella</em> infection, but do not affect cytokine expression or release. We observe that septin filaments assemble at the plasma membrane, and cleavage of GSDMD is impaired in septin-depleted cells. We found that septins regulate mitochondrial dynamics and the expression of NINJ1. Using a <em>Shigella</em>-zebrafish infection model, we show that septin-mediated pyroptosis is an <em>in vivo</em> mechanism of infection control. The discovery of septins as a mediator of pyroptosis may inspire innovative anti-bacterial and anti-inflammatory treatments.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1518-1528.e6"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2451945624003052/pdfft?md5=1fda9bdffaba472fc447836cdbe316e3&pid=1-s2.0-S2451945624003052-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141892170","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 : 2024-08-15DOI: 10.1016/j.chembiol.2024.07.005
Kanak Raina , Chris D. Forbes , Rebecca Stronk , Jonathan P. Rappi Jr. , Kyle J. Eastman , Nilesh Zaware , Xinheng Yu , Hao Li , Amit Bhardwaj , Samuel W. Gerritz , Mia Forgione , Abigail Hundt , Madeline P. King , Zoe M. Posner , Allison D. Correia , Andrew McGovern , David E. Puleo , Rebekka Chenard , James J. Mousseau , J. Ignacio Vergara , Craig M. Crews
We describe a protein proximity inducing therapeutic modality called Regulated Induced Proximity Targeting Chimeras or RIPTACs: heterobifunctional small molecules that elicit a stable ternary complex between a target protein (TP) selectively expressed in tumor cells and a pan-expressed protein essential for cell survival. The resulting co-operative protein-protein interaction (PPI) abrogates the function of the essential protein, thus leading to death selectively in cells expressing the TP. This approach leverages differentially expressed intracellular proteins as novel cancer targets, with the advantage of not requiring the target to be a disease driver. In this chemical biology study, we design RIPTACs that incorporate a ligand against a model TP connected via a linker to effector ligands such as JQ1 (BRD4) or BI2536 (PLK1) or CDK inhibitors such as TMX3013 or dinaciclib. RIPTACs accumulate selectively in cells expressing the HaloTag-FKBP target, form co-operative intracellular ternary complexes, and induce an anti-proliferative response in target-expressing cells.
{"title":"Regulated induced proximity targeting chimeras—RIPTACs—A heterobifunctional small molecule strategy for cancer selective therapies","authors":"Kanak Raina , Chris D. Forbes , Rebecca Stronk , Jonathan P. Rappi Jr. , Kyle J. Eastman , Nilesh Zaware , Xinheng Yu , Hao Li , Amit Bhardwaj , Samuel W. Gerritz , Mia Forgione , Abigail Hundt , Madeline P. King , Zoe M. Posner , Allison D. Correia , Andrew McGovern , David E. Puleo , Rebekka Chenard , James J. Mousseau , J. Ignacio Vergara , Craig M. Crews","doi":"10.1016/j.chembiol.2024.07.005","DOIUrl":"10.1016/j.chembiol.2024.07.005","url":null,"abstract":"<div><p>We describe a protein proximity inducing therapeutic modality called Regulated Induced Proximity Targeting Chimeras or RIPTACs: heterobifunctional small molecules that elicit a stable ternary complex between a target protein (TP) selectively expressed in tumor cells and a pan-expressed protein essential for cell survival. The resulting co-operative protein-protein interaction (PPI) abrogates the function of the essential protein, thus leading to death selectively in cells expressing the TP. This approach leverages differentially expressed intracellular proteins as novel cancer targets, with the advantage of not requiring the target to be a disease driver. In this chemical biology study, we design RIPTACs that incorporate a ligand against a model TP connected via a linker to effector ligands such as JQ1 (BRD4) or BI2536 (PLK1) or CDK inhibitors such as TMX3013 or dinaciclib. RIPTACs accumulate selectively in cells expressing the HaloTag-FKBP target, form co-operative intracellular ternary complexes, and induce an anti-proliferative response in target-expressing cells.</p></div>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"31 8","pages":"Pages 1490-1502.e42"},"PeriodicalIF":6.6,"publicationDate":"2024-08-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141899985","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}