Jakob Höppner,Tomasz Maciej Stepniewski,Thomas Dean,Miguel Diéguez-Eceolaza,Lauren My-Linh Tran,Samuel H. Gellman,Ross W. Cheloha,Asuka Inoue,Jana Selent,Harald Jüppner,Thomas J. Gardella
{"title":"Disease-causing helix-8 mutations reprogram parathyroid hormone receptor-1 effector selectivity","authors":"Jakob Höppner,Tomasz Maciej Stepniewski,Thomas Dean,Miguel Diéguez-Eceolaza,Lauren My-Linh Tran,Samuel H. Gellman,Ross W. Cheloha,Asuka Inoue,Jana Selent,Harald Jüppner,Thomas J. Gardella","doi":"10.1016/j.chembiol.2026.09.013","DOIUrl":"https://doi.org/10.1016/j.chembiol.2026.09.013","url":null,"abstract":"Helix-8 is an important determinant of G protein-coupled receptor (GPCR) effector selectivity, yet its functional relevance and contribution to human disease remain poorly understood. Two helix-8 mutations in parathyroid hormone receptor-1 (PTH1R), E4658.49K and E4698.53K, cause skeletal dysplasia, but their mechanistic effects on signaling were unclear. Using live-cell biosensors, structural modeling, and humanized knock-in mice, we show that these mutants reconfigure polar networks at the receptor’s cytoplasmic face to reshape effector coupling. E465K shifts PTH1R signaling away from Gαs toward Gαq activation, causing a switch in G protein selectivity. E469K stabilizes βarrestin engagement and enhances GRK2 recruitment, to thereby competitively limit G protein signaling. E469K-PTH1R mice show impaired calcemic responses to PTH, which are restored by a βarrestin-deficient PTH analog. These findings establish helix-8 as a critical determinant of GPCR effector selectivity and reveal mutation-driven rewiring of receptor signaling as a mechanism underlying PTH1R-associated disease.","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"2 1","pages":""},"PeriodicalIF":8.6,"publicationDate":"2026-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149151821","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}
Davide C Recchia, Richard Cardoso da Silva, Mathilde Noël, Douwe Ten Bulte, Ifigeneia Thomopoulou, Niklas Kupfer, Angela Topic, Sara Giuliani, Rodrigo Villaseñor, Tuncay Baubec
{"title":"A modular platform for in cellulo target engagement profiling of chromatin reader inhibitors.","authors":"Davide C Recchia, Richard Cardoso da Silva, Mathilde Noël, Douwe Ten Bulte, Ifigeneia Thomopoulou, Niklas Kupfer, Angela Topic, Sara Giuliani, Rodrigo Villaseñor, Tuncay Baubec","doi":"10.1016/j.chembiol.2026.09.009","DOIUrl":"https://doi.org/10.1016/j.chembiol.2026.09.009","url":null,"abstract":"<p><p>The dysregulation of acylation-reading bromodomain proteins is implicated in many diseases, making them important therapeutic targets. However, drug development is hindered by a lack of high-throughput cellular assays to measure target engagement and off-target binding in living cells. To address this gap, we developed a panel of cell lines that express synthetic bromodomain constructs, termed Acyl-eCRs. We demonstrate that Acyl-eCRs recapitulate the same response to bromodomain inhibitors and PROTACs as endogenous proteins, allowing for quantitative assessment of drug effects. We introduce two complementary flow cytometry-based assays to evaluate inhibitor-target engagement: a competitive binding assay leveraging PROTAC-induced degradation, and a nuclear retention assay measuring displacement of bromodomains from chromatin. Our approach circumvents laborious protein purification and in vitro characterization, providing a scalable, physiologically relevant method for assessing inhibitor potency and specificity. This platform provides a scalable strategy for quantitative target-engagement profiling of chromatin reader inhibitors in living cells.</p>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":" ","pages":""},"PeriodicalIF":9.0,"publicationDate":"2026-10-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149147151","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}
{"title":"Disrupting BRD4-BD1-p62 engagement restores protective autophagy","authors":"Jifa Zhang,Yu Cao,Chengyong Wu,Yang Li,Juncheng Chen,Doudou Wang,Yao Xiao,Binghan Xue,Cailing Gan,Xin Chen,Yuxi Wang,Tinghong Ye,Liang Ouyang","doi":"10.1016/j.chembiol.2026.09.007","DOIUrl":"https://doi.org/10.1016/j.chembiol.2026.09.007","url":null,"abstract":"Impaired protective autophagy contributes to idiopathic pulmonary fibrosis (IPF), yet therapeutic strategies that restore this process remain limited. Here, we identify SKLB-39b, a BRD4-BD1-selective inhibitor that restores protective autophagy and attenuates pulmonary fibrosis. SKLB-39b exhibits over 100-fold selectivity for BRD4-BD1 relative to BRD4-BD2 and nearly 10-fold greater affinity for BRD4-BD1 than for BRD2/3-BD1. Mechanistically, we identify a BRD4-BD1-p62 interaction that links BET bromodomain function to autophagy regulation. By engaging Leu92 and Ile146 through a hydrophobic binding mode, SKLB-39b disrupts this interaction, restores ULK1-dependent autophagy, and provides a structural framework for BD1-selective inhibitor design. SKLB-39b outperformed JQ-1 in suppressing fibroblast activation, epithelial-mesenchymal transition, and collagen deposition in experimental fibrosis while showing favorable pharmacokinetic exposure and no overt toxicity in the mouse studies performed. These findings establish the BRD4-BD1-p62 interface as a druggable target for restoring protective autophagy in IPF.","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"25 1","pages":""},"PeriodicalIF":8.6,"publicationDate":"2026-10-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149124077","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}
{"title":"Targeting ZDHHC15-mediated S-acylation of CD155 reactivates anti-tumor immunity","authors":"Xuanpu Zhang,Jie Zou,Jian Lin,Yu Xiao,Yue Zhu,Bokan Chen,Ziye Zhu,Yizhen Ding,Xinmei Zhou,Yinghong Yang,Xiaoyi Wang,Weijie Zhang,Min Zhang,Xia Lin,Pinglong Xu,Baoen Chen","doi":"10.1016/j.chembiol.2026.09.006","DOIUrl":"https://doi.org/10.1016/j.chembiol.2026.09.006","url":null,"abstract":"The immune checkpoint CD155 is frequently upregulated across multiple malignancies and drives tumor progression and immune evasion, yet the posttranslational mechanisms regulating its stability and activity remain unclear. Here, we identify S-acylation as a key regulator of CD155 stability and immunosuppressive function. Using chemical reporters, we demonstrate that CD155 is S-acylated at three cytoplasmic cysteines by the acyltransferase ZDHHC15 and deacylated by the thioesterases PPT1/APT1. S-acylation protects CD155 from lysosomal degradation and promotes its interaction with Sprouty2 to sustain oncogenic ERK signaling. Genetic inhibition of ZDHHC15 disrupts CD155 S-acylation, triggering CD155 degradation, enhancing CD8+ T cell infiltration, and suppressing tumor growth in syngeneic models. We further show that the FDA-approved CDK4/6 inhibitor abemaciclib promotes CD155 degradation by upregulating PPT1 and enhancing lysosome biogenesis. Our work identifies reversible CD155 S-acylation as a druggable regulatory mechanism that links protein lipidation to immune checkpoint stability and reveals an unexpected immune-potentiating mechanism of abemaciclib.","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"3 1","pages":""},"PeriodicalIF":8.6,"publicationDate":"2026-10-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149090229","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}
{"title":"Rational design of protease-resistant lipopeptides for combating antibiotic-resistant bacteria.","authors":"Jiajun Wang, Haoran Xu, Yu Sun, Na Dong, Weikang Yu, Shiqi He, Anshan Shan","doi":"10.1016/j.chembiol.2026.09.001","DOIUrl":"https://doi.org/10.1016/j.chembiol.2026.09.001","url":null,"abstract":"<p><p>Given the surge in antibiotic resistance, antimicrobial peptides have emerged as promising alternatives to conventional antibiotics; however, their therapeutic potential remains hindered by proteolytic instability and insufficient potency. Herein, we integrated an anti-proteolytic Arg-Pro scaffold with N-terminal fatty acid modification to improve peptide stability and membrane-binding affinity. A series of lipopeptides was synthesized and systematically evaluated for antimicrobial efficacy, stability, and cytotoxicity, yielding C<sub>12</sub>RP7 as the lead candidate. C<sub>12</sub>RP7 exhibited potent broad-spectrum activity against 20 tested strains, including multi-drug resistant pathogens, with a high therapeutic index (TI = 64.20) and low toxicity. It also retained functional stability in the presence of physiological salts, serum, and proteases. In murine models of peritonitis and enteritis, C<sub>12</sub>RP7 treatment significantly reduced bacterial burden, attenuated systemic inflammation, restored intestinal barrier integrity, and modulated the gut microbiota. These findings support C<sub>12</sub>RP7 as a protease-resistant, potent, and biocompatible lipopeptide with potential for treating bacterial infections.</p>","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":" ","pages":""},"PeriodicalIF":9.0,"publicationDate":"2026-09-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149101839","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}
Christina E. Lee,Judith Martinez,Anna Amine,Wei Zhang,Chih-Hao Lu,Xingyuan Zhang,He You,Ching-Ting Tsai,Melissa L. Nakamoto,Bianxiao Cui
{"title":"Curved adhesions resist disruption by high-affinity αv-integrin inhibitors","authors":"Christina E. Lee,Judith Martinez,Anna Amine,Wei Zhang,Chih-Hao Lu,Xingyuan Zhang,He You,Ching-Ting Tsai,Melissa L. Nakamoto,Bianxiao Cui","doi":"10.1016/j.chembiol.2026.08.017","DOIUrl":"https://doi.org/10.1016/j.chembiol.2026.08.017","url":null,"abstract":"Integrins are transmembrane adhesion receptors and major therapeutic targets in cancer and fibrotic diseases. High-affinity αv-integrin inhibitors, including cilengitide and MK-0429, were developed to block integrin-dependent cell adhesions. These inhibitors have been shown to disrupt integrin-mediated focal adhesions on flat, ligand-coated surfaces. Here, we demonstrate that these inhibitors are ineffective against curved adhesions, a class of integrin-mediated adhesion structures that assemble in response to membrane curvature. We find that, unlike focal adhesions, curved adhesions form under low mechanical tension, rendering them resistant to these inhibitors that act by blocking force transmission. As a result, these inhibitors also do not prevent cancer cell invasion into soft, fibrous three-dimensional extracellular matrices, a process that depends on curved adhesions. These findings expose a limitation of current integrin-targeting strategies and may help explain why some inhibitors have underperformed in clinical trials.","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"53 1","pages":""},"PeriodicalIF":8.6,"publicationDate":"2026-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149067374","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}
Cinzia Barresi,Eleonora Maino,Weiqiang Chen,Jakob J. Prömer,Alexia Bock-Pereda,Markus Hartl,Markus A. Rüegg,Ruth Herbst
{"title":"Crk/Crk-L-mediated Rapgef1 recruitment couples MuSK signaling to AChR anchoring at the neuromuscular junction","authors":"Cinzia Barresi,Eleonora Maino,Weiqiang Chen,Jakob J. Prömer,Alexia Bock-Pereda,Markus Hartl,Markus A. Rüegg,Ruth Herbst","doi":"10.1016/j.chembiol.2026.08.014","DOIUrl":"https://doi.org/10.1016/j.chembiol.2026.08.014","url":null,"abstract":"The neuromuscular junction (NMJ) is established through reciprocal signaling between motoneurons and skeletal muscle, a process critically dependent on the receptor tyrosine kinase MuSK. Although MuSK activation by Agrin, Lrp4, and Dok-7 is well characterized, the temporal organization of downstream signaling remains unclear. Here, we define the MuSK proximity proteome using TurboID-based proximity labeling combined with mass spectrometry. We identified 74 MuSK-associated proteins, including known substrates and previously unreported interactors. Among early signaling components, the adapter Crk-L emerged as a central hub linking MuSK to small GTPase pathways. We further identified Rapgef1 as a Crk-L-associated effector required for Agrin-dependent acetylcholine receptor (AChR) phosphorylation and clustering. Rapgef1 loss in adult muscle led to NMJ fragmentation. We propose that these defects result from reduced cytoskeletal anchorage of AChRs due to compromised Rapsyn/AChR interaction. These findings uncover Rapgef1 as a key regulator of MuSK-mediated signaling and highlight dynamic mechanisms governing NMJ formation and maintenance.","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"88 1","pages":""},"PeriodicalIF":8.6,"publicationDate":"2026-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149013603","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}
Signe Mathiasen,Jozsef Meszaros,Chase Webb,Peter Geggier,Wesley B. Asher,Nevin A. Lambert,Jonathan A. Javitch
{"title":"G protein diffusion states reveal receptor binding dynamics at single-molecule scale","authors":"Signe Mathiasen,Jozsef Meszaros,Chase Webb,Peter Geggier,Wesley B. Asher,Nevin A. Lambert,Jonathan A. Javitch","doi":"10.1016/j.chembiol.2026.08.013","DOIUrl":"https://doi.org/10.1016/j.chembiol.2026.08.013","url":null,"abstract":"G protein-coupled receptors (GPCRs) signal through heterotrimeric G proteins, yet the kinetics of receptor-G protein interactions in cells remain poorly understood. Using single-molecule imaging, we tracked individual Gαi1 proteins diffusing in the plasma membrane, expecting that receptor binding should slow these rapidly diffusing lipid-anchored proteins to match the substantially slower transmembrane GPCRs. Gαi1-Halo showed no detectable slowing upon dopamine D2 receptor activation. In contrast, the nucleotide-decoupled Gαi1 4A-Halo mutant exhibited receptor-dependent slowing with a lifetime of ∼0.8 s. Agonist increased the Gαi1 4A-Halo association rate without affecting the lifetime of the receptor-engaged complex, consistent with the idea that agonist efficacy is reflected in an increased probability of productive G protein engagement. These findings suggest that native GPCR-G protein interactions are not resolved at 15 ms time resolution and establish a diffusion-based approach for studying receptor-G protein kinetics in living cells.","PeriodicalId":265,"journal":{"name":"Cell Chemical Biology","volume":"30 1","pages":""},"PeriodicalIF":8.6,"publicationDate":"2026-09-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"149010173","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}
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