The FASEB Journal最新文献

{"title":"ZUP1 is a key component of the MAVS complex and acts as a protector of host against viral invasion","authors":"Wenyan Hao,&nbsp;Mengfan Guo,&nbsp;Xin Ji,&nbsp;Xuyang Zhao,&nbsp;Minglu Zhu,&nbsp;Yan Jin,&nbsp;Guangxi Wang,&nbsp;Jiawen Feng,&nbsp;Dan Lu,&nbsp;Yuxin Yin","doi":"10.1096/fj.202401661RRR","DOIUrl":"https://doi.org/10.1096/fj.202401661RRR","url":null,"abstract":"<p>Zinc finger-containing ubiquitin peptidase 1 (ZUP1) is a protein characterized by four N-terminal zinc finger domains and a C-terminal deubiquitinase (DUB) domain. While it is associated with the DNA damage response, the role of ZUP1 in innate immunity remains unclear. Here, we identify ZUP1 as a crucial component of the mitochondrial antiviral signaling (MAVS) complex, essential for host antiviral defense. We show that viral infection significantly upregulates ZUP1 expression, and mice lacking ZUP1 exhibit impaired type I interferon (IFN) production and increased susceptibility to viral infection, as evidenced by higher mortality rates. This underscores the protective role of ZUP1 in host immunity. Mechanistically, ZUP1 binds to MAVS through its C-terminal domain independently of DUB activity. Instead, ZUP1 utilizes its zinc finger domains, particularly the third zinc finger, to directly bind viral RNA. This interaction enhances the association of ZUP1 with MAVS and promotes its aggregation on mitochondria during viral infection. ZUP1 also interacts with TBK1 and NEMO within the MAVS complex, facilitating IRF3 activation and type I IFN production. These findings establish ZUP1 as a zinc finger-containing regulator that amplifies MAVS-dependent antiviral immunity, linking viral RNA recognition to downstream signaling and highlighting potential targets for therapeutic intervention against viral infections.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143632927","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":"Research progress on the anti-aging effect of polysaccharides of traditional Chinese medicine: Using Caenorhabditis elegans as an animal model","authors":"Jiazi Lin,&nbsp;Jiamin Yu,&nbsp;Xiao Wang,&nbsp;Ruixiang Shi,&nbsp;Yefang Liang,&nbsp;Jianhua Li,&nbsp;Tong Zhou,&nbsp;Chengkai Chen,&nbsp;Xiaodong Duan,&nbsp;Yongan Deng,&nbsp;Simin Yang,&nbsp;Shuting Zeng,&nbsp;Xuejuan Shen,&nbsp;Xiangyu Chen,&nbsp;Yi Wang,&nbsp;Guibo Sun,&nbsp;Zunpeng Shu","doi":"10.1096/fj.202403250RR","DOIUrl":"https://doi.org/10.1096/fj.202403250RR","url":null,"abstract":"<p>With the growing elderly population and increasing incidence of various aging-related diseases, the scientific community is faced with an urgent challenge to identify natural anti-aging agents. Traditional Chinese medicine (TCM) polysaccharides have been proven to have good anti-aging activities. This article reviews the literature on the anti-aging pathways of traditional Chinese medicine polysaccharides applied to <i>Caenorhabditis elegans</i> models in the past decade. In our study, we found that 45 TCM polysaccharides from 28 genera and 26 families could delay the aging process of <i>C. elegans</i>. Traditional Chinese medicine polysaccharides delay the aging of <i>C. elegans</i> mainly by anti-oxidative stress, eliminating free radicals, repairing DNA damage, and insulin/insulin-like growth factor signaling pathway (IIS signaling pathway). In addition, an increasing number of traditional Chinese medicine polysaccharides have been found to prolong the lifespan of <i>C. elegans</i> by reducing inflammation, regulating intestinal flora, and affecting immune cell function. In this paper, <i>C. elegans</i> was used as an animal model to clarify the anti-aging pathway of traditional Chinese medicine polysaccharides, so as to provide theoretical guidance for future research and clinical experiments on the anti-aging effect of traditional Chinese medicine polysaccharides.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202403250RR","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143612482","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":"HuR inhibition reduces post-ischemic cardiac remodeling by dampening myocyte-dependent inflammatory gene expression and the innate immune response","authors":"Samuel Slone,&nbsp;Sarah R. Anthony,&nbsp;Lisa C. Green,&nbsp;Sharon Parkins,&nbsp;Pooja Acharya,&nbsp;Daniel A. Kasprovic,&nbsp;Kelsi Reynolds,&nbsp;Robert M. Jaggers,&nbsp;Michelle L. Nieman,&nbsp;Perwez Alam,&nbsp;Xiaoqing Wu,&nbsp;Sudeshna Roy,&nbsp;Jeffrey Aubé,&nbsp;Liang Xu,&nbsp;Zihai Li,&nbsp;John N. Lorenz,&nbsp;A. Phillip Owens,&nbsp;Onur Kanisicak,&nbsp;Michael Tranter","doi":"10.1096/fj.202400532RRR","DOIUrl":"https://doi.org/10.1096/fj.202400532RRR","url":null,"abstract":"<p>The RNA-binding protein human antigen R (HuR) has been shown to reduce cardiac remodeling following both myocardial infarction and cardiac pressure overload, but the full extent of the HuR-dependent mechanisms within cells of the myocardium has yet to be elucidated. Wild-type mice were subjected to 30 min of cardiac ischemia (via LAD occlusion) and treated with a novel small molecule inhibitor of HuR at the time of reperfusion, followed by direct in vivo assessment of cardiac structure and function. Direct assessment of HuR-dependent mechanisms was done in vitro using neonatal rat ventricular myocytes (NRVMs) and bone marrow-derived macrophages (BMDMs). HuR activity is increased within 2 h after ischemia/reperfusion (I/R) and is necessary for early post-I/R inflammatory gene expression in the myocardium. Despite an early reduction in inflammatory gene expression, HuR inhibition has no effect on initial infarct size at 24 h post-I/R. However, pathological remodeling is reduced with preserved cardiac function at 2 weeks post-I/R upon HuR inhibition. RNA sequencing analysis of gene expression in NRVMs treated with LPS to model damage-associated molecular pattern (DAMP)-mediated activation of toll-like receptors (TLRs) demonstrates a HuR-dependent regulation of pro-inflammatory chemokine and cytokine gene expression in cardiomyocytes. Importantly, we show that conditioned media transfer from NRVMs pre-treated with HuR inhibitor loses the ability to induce inflammatory gene expression and M1-like polarization in bone marrow-derived macrophages (BMDMs) compared to NRVMs treated with LPS alone. Functionally, HuR inhibition reduces macrophage infiltration to the post-ischemic myocardium in vivo. Additionally, we show that LPS-treated NRVMs induce the migration of peripheral blood monocytes in a HuR-dependent endocrine manner. These studies demonstrate that HuR is necessary for early pro-inflammatory gene expression in cardiomyocytes following I/R injury that subsequently mediates monocyte recruitment and macrophage activation in the post-ischemic myocardium.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202400532RRR","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143612562","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":"Muscle-specific Keap1 deletion enhances force production but does not prevent inactivity-induced muscle atrophy in mice","authors":"Edwin R. Miranda,&nbsp;Justin L. Shahtout,&nbsp;Shinya Watanabe,&nbsp;Norah Milam,&nbsp;Takuya Karasawa,&nbsp;Subhasmita Rout,&nbsp;Donald L. Atkinson,&nbsp;William L. Holland,&nbsp;Micah J. Drummond,&nbsp;Katsuhiko Funai","doi":"10.1096/fj.202402810R","DOIUrl":"https://doi.org/10.1096/fj.202402810R","url":null,"abstract":"<p>Immobilization-associated muscle atrophy and weakness appear to be driven in part by oxidative stress. Nuclear Factor Erythroid 2-Related Factor 2 (NRF2) is a critical redox rheostat that regulates oxidative stress responses, and its deletion is known to accelerate muscle atrophy and weakness during aging (sarcopenia) or denervation. Conversely, pharmacologic activation of NRF2 extends mouse lifespan and attenuates sarcopenia. Similarly, deletion of Kelch-like ECH-associated Protein 1 (Keap1), a negative regulator of NRF2, enhances exercise capacity. The purpose of this study was to determine whether muscle-specific Keap1 deletion is sufficient to prevent muscle atrophy and weakness in mice following 7 days of hindlimb unloading (HU). To test this hypothesis, control (Ctrl) and tamoxifen-inducible, muscle-specific Keap1 knockout (mKO) mice were subjected to either normal housing (Sham) or HU for 7 days. Activation of NRF2 in muscle was confirmed by increased mRNA of NRF2 targets thioredoxin 1 (Txn1) and NAD(P)H quinone dehydrogenase 1 (NQO1) in mKO mice. Keap1 deletion had an effect to increase force-generating capacity at baseline. However, muscle masses, cross-sectional area, and ex vivo force were not different between mKO and Ctrl HU mice. In addition, muscle 4-hydroxynonenal-modified proteins and protein carbonyls were unaffected by Keap1 deletion. These data suggest that NRF2 activation improves muscle force production during ambulatory conditions but is not sufficient to prevent muscle atrophy or weakness following 7 days of HU.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202402810R","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143612509","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":"Prebiotics pectin and resistant starch-type 4 stimulate peptide YY and cholecystokinin to promote satiety, and improve gut microbiota composition","authors":"Souvik Patra,&nbsp;Savana L. Everhart Nunn,&nbsp;Gizem Levent,&nbsp;Prasanth K. Chelikani","doi":"10.1096/fj.202403239R","DOIUrl":"https://doi.org/10.1096/fj.202403239R","url":null,"abstract":"<p>Dietary prebiotics pectin and resistant starch type-4 (RS-4) promote satiety and alter gut microbiota; however, the underlying neurohormonal mechanisms of satiety remain poorly understood. We determined the effects of pectin, RS-4, and their combination on energy balance and gut microbiota composition, and assessed whether the gut hormones peptide YY (PYY) and cholecystokinin (CCK) play a role in fiber-induced satiety. High-fat diet –induced obese male rats (<i>n</i> = 7–8/group) were fed either control, pectin, RS-4, or a combination of pectin and RS-4 diet. We found that pectin, RS-4, and their combination decreased food intake. Pectin alone, or combined with RS-4, shifted substrate utilization towards fat and reduced gains in weight and adiposity. Pectin alone or combined with RS-4 enhanced the expression and plasma concentrations of PYY and CCK. Importantly, systemic blockade of PYY-Y2 and CCK-1 receptors attenuated the hypophagic effects of pectin, and CCK-1 receptor blockade partly attenuated the hypophagia from RS-4. The prebiotics significantly altered fecal β-diversity metrics, suggestive of improvements in gut microbiota composition. Pectin and RS-4 alone, or in combination, were associated with increased relative abundance of phylum <i>Bacteroidota</i>, decreased <i>Firmicutes</i>, and increased concentrations of amino acids and biogenic amines in feces. Collectively, these findings suggest that dietary pectin and RS-4 improved energy balance and gut microbiota composition, and importantly, demonstrated that the satiety effects of these diets were mediated, in part, via enhanced endogenous PYY and CCK signaling.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143612563","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":"Exploring the impact of RPN1 on tumorigenesis and immune response in cancer","authors":"Pengfei Luo,&nbsp;Zhimin Li,&nbsp;Haodong He,&nbsp;Yuanbin Tang,&nbsp;Lijun Zeng,&nbsp;Lunqi Luo,&nbsp;Lianjie Ouyang,&nbsp;Meiling Wen,&nbsp;Yuehua Li,&nbsp;Yongjun Jiang","doi":"10.1096/fj.202401088R","DOIUrl":"https://doi.org/10.1096/fj.202401088R","url":null,"abstract":"<p>The ribophorin family, including RPN1, has been associated with tumor progression, but its specific role in pan-cancer dynamics remains unclear. Using data from TCGA, GTEx, and Ualcan databases, we investigated the relationship of RPN1 with prognosis, genomic alterations, and epigenetic modifications across various cancers. Differential analysis revealed elevated RPN1 expression in multiple cancer types, indicating a potential prognostic value. Amplification was the predominant mutation type of RPN1 in pan-cancer, with notable correlations with DNA methylation and copy number variation. Gene set variation analysis identified RPN1's involvement in cancer development, immunity, and metabolism. Additionally, RPN1 expression correlated with the tumor microenvironment, immune response factors, and response to anti-tumor therapies. Functional validation in triple-negative breast cancer, glioblastoma, and bladder cancer cell lines demonstrated the role of RPN1 in tumor cell proliferation and migration. Our findings highlight RPN1 as a potential biomarker for cancer diagnosis and treatment response in pan-cancer therapy.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143602742","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":"Controversy and multiple roles of the solitary nucleus receptor Nur77 in disease and physiology","authors":"Yanteng Wang,&nbsp;Na Li,&nbsp;Wenwei Guan,&nbsp;Difei Wang","doi":"10.1096/fj.202402775RR","DOIUrl":"https://doi.org/10.1096/fj.202402775RR","url":null,"abstract":"<p>Neuron-derived clone 77 (Nur77), a member of the orphan nuclear receptor family, is expressed and activated rapidly in response to diverse physiological and pathological stimuli. It exerts complex biological functions, including roles in the nervous system, genome integrity, cell differentiation, homeostasis, oxidative stress, autophagy, aging, and infection. Recent studies suggest that Nur77 agonists alleviate symptoms of neurodegenerative diseases, highlighting its potential as a therapeutic target in such conditions. In cancer, Nur77 demonstrates dual roles, acting as both a tumor suppressor and promoter, depending on the cancer type and stage, making it a controversial yet promising anticancer target. This review provides a structured analysis of the functions of Nur77, focusing on its physiological and pathological roles, therapeutic potential, and existing controversies. Emphasis is placed on its emerging applications in neurodegenerative diseases and cancer, offering key insights for future research and clinical translation.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202402775RR","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143602840","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":"Potential biological roles of exosomal non-coding RNAs in breast cancer","authors":"Xiang Li,&nbsp;Junyi Gong,&nbsp;Xiang Ni,&nbsp;Junli Yin,&nbsp;Yi Zhang,&nbsp;Zheng Lv","doi":"10.1096/fj.202500022R","DOIUrl":"https://doi.org/10.1096/fj.202500022R","url":null,"abstract":"<p>Breast cancer (BC) is one of the most common malignant tumors among women, accounting for 24.5% of all cancer cases and leading to 15.5% of cancer-related mortality. The treatment of BC patients remains a significant challenge due to the disease's high invasiveness, elevated metastatic potential, substantial drug resistance, and high recurrence rate. Exosomes, which are lipid-bilayer extracellular vesicles ranging in size from 30 to 150 nm, mediate intercellular communication between tumor cells and surrounding cells in the tumor microenvironment by transferring various bioactive substances, such as proteins, lipids, and nucleic acids. Recently, growing evidence has demonstrated that non-coding RNAs (ncRNAs) are enriched in exosomes and play a critical role in regulating cell proliferation, metastasis, drug resistance, and angiogenesis in BC. Consequently, exosomal ncRNAs have emerged as a promising therapeutic target for BC treatment, given their involvement in multiple processes of cancer progression. This review provides a comprehensive and in-depth analysis of emerging exosomal ncRNAs in BC, highlighting their potential biological mechanisms and advanced applications in BC treatment.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1096/fj.202500022R","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143602464","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":"Spermidine alleviates copper-induced oxidative stress, inflammation and cuproptosis in the liver","authors":"Xin Wang,&nbsp;Weikang Ling,&nbsp;Yang Zhu,&nbsp;Chengweng Ji,&nbsp;Xiaoguang An,&nbsp;Yuxin Qi,&nbsp;Shuo Li,&nbsp;Chengye Zhang,&nbsp;Ruixue Tong,&nbsp;Dongmei Jiang,&nbsp;Bo Kang","doi":"10.1096/fj.202403002R","DOIUrl":"https://doi.org/10.1096/fj.202403002R","url":null,"abstract":"<p>Copper exposure poses potential detrimental effects on both public and ecosystem health. Spermidine, an antioxidant, has shown promise in reducing oxidative stress and inflammation within the liver. However, its specific role in mitigating copper-induced hepatic cuproptosis and disturbances in copper metabolism remains unexplored. Consequently, this research aims to investigate to examine the impact of spermidine on hepatic cuproptosis and the related disturbances in copper metabolism. In the study, we established a model of copper-induced liver toxicity by feeding C57BL/6 mice a high-copper diet for three months. Histopathological and biochemical analyses revealed that copper exposure induced hepatic inflammatory cell infiltration, hepatocyte degeneration, elevated levels of MDA, ROS, and Cu²⁺ accumulation in the liver, and increased ALT and AST activities in serum (<i>p</i> &lt; .05). Regarding inflammation, copper exposure significantly increased serum levels of IL-1β, IL-6, and TNF-α (<i>p</i> &lt; .05), upregulated <i>TNF-α</i> and <i>IFN-γ</i> expression, and downregulated <i>IL-10</i> expression in the liver (<i>p</i> &lt; .05). Meanwhile, copper exposure inhibited the expression of copper metabolism and Fe-S cluster-related proteins (<i>p</i> &lt; .05). Exogenous spermidine administration effectively reduced ROS, MDA, and Cu²⁺ accumulation in the liver, while also decreasing ALT and AST activites, IL-1β, IL-6, and TNF-α levels in the serum (<i>p</i> &lt; .05), and downregulated <i>TNF-α</i> and <i>IFN-γ</i> expression (<i>p</i> &lt; .001). Additionally, spermidine combined with CuSO₄ treatment significantly promotes the expression of copper metabolism and Fe-S cluster-related proteins, compared to the CuSO₄ group (<i>p</i> &lt; .05). In summary, spermidine reduces Cu²⁺ accumulation in the liver, alleviates hepatic cuproptosis, oxidative damage, and inflammation, and exerts a protective effect on the liver.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143602841","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":"Investigation of structural and dynamic properties of the Butyrophilin BTN3A1/BTN2A1 cytoplasmic complex by 19F solution NMR","authors":"Khiem Nguyen,&nbsp;Chia-Hung Christine Hsiao,&nbsp;Yiming Jin,&nbsp;Andrew J. Wiemer,&nbsp;Olga Vinogradova","doi":"10.1096/fj.202402975","DOIUrl":"https://doi.org/10.1096/fj.202402975","url":null,"abstract":"<p>Butyrophilin 3A1 (BTN3A1) is an integral membrane protein capable of detecting phosphoantigens, like (<i>E</i>)-4-hydroxy-3-methyl-but-2-enyl diphosphate (HMBPP), through its internal B30.2 domain. Detection of phosphoantigens leads to interactions with butyrophilin 2A1 and the subsequent activation of γδ-T cells. Though crystallography and functional assays have been crucial for determining vital residues of the BTN3A1/HMBPP/BTN2A1 complex, the mechanism for signal transduction is still unclear. Here, we utilize ¹⁹F solution NMR to observe potential conformational and dynamic changes of specific residues upon complex formation. With point mutants of BTN3A1, we show that W421C, T449C, and T506C are residues that are influenced by HMBPP and BTN2A1 association, while T304C, G323C, C387, and C511 are not impacted. ¹⁹F labeling of W421C reduces the binding affinity of BTN2A1 toward BTN3A1/HMBPP, which indicates that W421 is located at the binding interface. T506 is located away from the phosphoantigen binding site, so its observable chemical shift perturbation suggests that there is a larger conformational change of the BTN3A1 B30.2 domain upon binding HMBPP and BTN2A1. The juxtamembrane residues, T304C, and G323C are not affected, showing that the changes are localized within the B30.2 domain of BTN3A1. Using BTN3A1 T449C, we were able to detect differential binding modes of synthetic HMBPP analogs, showing that it is possible to assess differences in protein conformations that are induced by binding of different ligands. Taken together, these findings illustrate the dynamic processes involved in phosphoantigen detection by the HMBPP receptor.</p>","PeriodicalId":50455,"journal":{"name":"The FASEB Journal","volume":"39 6","pages":""},"PeriodicalIF":4.4,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143602740","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}