{"title":"Theileria annulata Hijacks Host Signaling: Integrated Phosphoproteomics and transcriptomics Unveils ERK1/2 as a Central Regulator of Host Transcription Factors.","authors":"Debabrata Dandasena, Vengatachala Moorthy A, Akash Suresh, Vasundhra Bhandari, Sonti Roy, Paresh Sharma","doi":"10.1016/j.mcpro.2025.100992","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100992","url":null,"abstract":"<p><p>THEILERIA: transformed bovine leukocytes exhibit cancer-like characteristics, but the molecular mechanisms driving these transformations remain unclear. This study provides the first comprehensive phosphoproteomic analysis of both host and parasite in Theileria annulata-infected leukocyte cell lines. We show that T. annulata significantly induces changes in the host protein phosphorylation, impacting key cancer-related processes such as apoptosis suppression, CAMK signaling, and telomere maintenance. A pivotal finding is the parasite's manipulation of the MAPK pathway via sustained ERK1/2 activation, which regulates the phosphorylation of critical transcription factors like RUNX3, FOSL2, BCL6, c-JUN, JUNB, and c-MYC. Transcriptomic analysis of genes controlled by these transcription factors confirmed their role in T. annulata replication. ERK inhibition disrupts phosphorylation, deactivates these transcription factors, and induces apoptosis in infected cells. This underscores the ERK-AP-1 axis as a central mechanism of Theileria pathogenesis and a promising therapeutic target. Additionally, parasite-specific phosphoproteins and kinases were identified, offering new insights into therapeutic strategies to combat infection.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100992"},"PeriodicalIF":6.1,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144078705","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}
Nina A Khristenko, Konstantin O Nagornov, Camille Garcia, Natalia Gasilova, Megan Gant, Karen Druart, Anton N Kozhinov, Laure Menin, Julia Chamot-Rooke, Yury O Tsybin
{"title":"Top-Down and Middle-Down Mass Spectrometry of Antibodies.","authors":"Nina A Khristenko, Konstantin O Nagornov, Camille Garcia, Natalia Gasilova, Megan Gant, Karen Druart, Anton N Kozhinov, Laure Menin, Julia Chamot-Rooke, Yury O Tsybin","doi":"10.1016/j.mcpro.2025.100989","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100989","url":null,"abstract":"<p><p>Therapeutic antibodies, primarily immunoglobulin G-based monoclonal antibodies, are developed to treat cancer, autoimmune disorders, and infectious diseases. Their large size, structural complexity, and heterogeneity pose significant analytical challenges, requiring the use of advanced characterization techniques. This review traces the 30-year evolution of top-down (TD) and middle-down (MD) mass spectrometry (MS) for antibody analysis, beginning with their initial applications and highlighting key advances and challenges throughout this period. TD MS allows for the analysis of intact antibodies, and MD MS performs analysis of the antibody subunits, even in complex biological samples. Both approaches preserve critical quality attributes such as sequence integrity, post-translational modifications (PTMs), disulfide bonds, and glycosylation patterns. Key milestones in TD and MD MS of antibodies include the use of structure-specific enzymes for subunit generation, the implementation of high-resolution mass spectrometers, and the adoption of non-ergodic ion activation methods such as electron transfer dissociation (ETD), electron capture dissociation (ECD), ultraviolet photodissociation (UVPD), and matrix-assisted laser desorption/ionization in-source decay (MALDI-ISD). The combination of complementary dissociation methods and the use of consecutive ion activation approaches has further enhanced TD/MD MS performance. The current TD MS record of antibody sequencing with terminal product ions is about 60% sequence coverage obtained using the activated ion-ETD approach on a high-resolution MS platform. Current MD MS analyses with about 95% sequence coverage were achieved using combinations of ion activation and dissociation techniques. The review explores TD and MD MS analysis of novel mAb modalities, including antibody-drug conjugates, bispecific antibodies, and endogenous antibodies from biofluids as well as immunoglobulin A and M-type classes. Content.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100989"},"PeriodicalIF":6.1,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144078715","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":"Temporal wheat proteome remodeling by deoxynivalenol reveals novel detoxification signatures and strategies across cultivars.","authors":"Reid Buchanan, Kholoud Shaban, Boyan Liu, Norris Chan, Mitra Serajazari, Jennifer Geddes-McAlister","doi":"10.1016/j.mcpro.2025.100988","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100988","url":null,"abstract":"<p><p>Fusarium head blight (FHB) is a globally devastating fungal disease resulting in reduced grain yield and quality, along with contamination of grains with dangerous mycotoxins. Consumption of such mycotoxins by humans through processed food or livestock through feed has downstream implications for human and animal health. This interconnectivity across the environment, animal, and human health defines the One Health problem of threatened food safety and security. In this study, we explore remodeling of the wheat proteome upon exposure to a common mycotoxin, deoxynivalenol (DON). We investigate cultivar-specific responses to DON exposure in FHB-susceptible (Norwell) and -resistant (Sumai#3) cultivars across a continuum of exposure (i.e., 24 and 120 hours post inoculation), and upon low (i.e., 0.1 mg/mL) and high (1.0 mg/mL) levels of the mycotoxin. This complex experimental design enables us to tease apart the dynamic relationship between each cultivar and DON tolerance. Specifically, we define precise proteins and broad categories of remodeling that are common (i.e., reduction in photosynthesis) and exclusive (i.e., glycosyltransferase) to the cultivars and align with anticipated protective mechanisms. Moreover, we adapted an in vitro DON tolerance expression system and determined that induction of an ubiquinol oxidase (UniProt ID: A0A3B6B5K8) provides heightened protection for yeast growth relative to the negative control, as well as increased protection compared to a well-defined DON detoxifying protein. Our study suggests a new avenue for identification and characterization of novel DON detoxifying proteins as putative biomarkers for selected breeding strategies. Such strategies support the production of wheat varieties with increased tolerance to DON for improved global food safety and security.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100988"},"PeriodicalIF":6.1,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144007943","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}
Keaton L Mertz, Jordahl, Colin A Hemme, Mitchell D Probasco, Dylan S Forbes, Peter L Ducos, Austin Z Salome, Michael S Westphall, Scott T Quarmby, Timothy Grant, Joshua J Coon
{"title":"Laser-Induced Rehydration of Cryo-Landed Proteins Restores Native Structure.","authors":"Keaton L Mertz, Jordahl, Colin A Hemme, Mitchell D Probasco, Dylan S Forbes, Peter L Ducos, Austin Z Salome, Michael S Westphall, Scott T Quarmby, Timothy Grant, Joshua J Coon","doi":"10.1016/j.mcpro.2025.100987","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100987","url":null,"abstract":"<p><p>The use of native mass spectrometry (MS) to land biological molecules for subsequent cryogenic electron microscopy (cryoEM) imaging and three-dimensional reconstruction has gained momentum in recent years as a means to overcome longstanding challenges posed by traditional cryoEM sample preparation. However, recent results obtained with this approach have been constrained by low resolution and the compaction of cryo-landed particles, likely due to dehydration during exposure to vacuum. Here, we describe a new sample preparation method that uses a laser integrated into a cryogenic soft-landing apparatus to liquefy precisely deposited amorphous ice, rehydrating particles and restoring their solution structure prior to rapid revitrification via the thermal mass of the grid. With this technique, we demonstrate the reconstruction of cryo-landed, rehydrated, and revitrified β-galactosidase that is comparable in resolution to that achieved with plunge freezing. Further, these particles are not compacted, matching the known structure and conformation obtained with traditionally plunge-frozen particles. These results establish the viability of coupling native MS with cryoEM for high-resolution structural determination without the limitations imposed by conventional sample preparation, and they open a path to solving previously inaccessible molecules and to integrating MS capabilities such as gas-phase purification to complex samples such as cell lysates.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100987"},"PeriodicalIF":6.1,"publicationDate":"2025-05-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144010171","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}
Shi Yan, Jorick Vanbeselaere, Callum Ives, David Stenitzer, Lena Nuschy, Florian Wöls, Katharina Paschinger, Elisa Fadda, Johannes Stadlmann, Iain B H Wilson
{"title":"Glycoproteomic and single-protein glycomic analyses reveal zwitterionic N-glycans on natural and recombinant proteins derived from insect cells.","authors":"Shi Yan, Jorick Vanbeselaere, Callum Ives, David Stenitzer, Lena Nuschy, Florian Wöls, Katharina Paschinger, Elisa Fadda, Johannes Stadlmann, Iain B H Wilson","doi":"10.1016/j.mcpro.2025.100981","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100981","url":null,"abstract":"<p><p>Insect cells are a convenient cell factory to produce recombinant glycoproteins. Their glycosylation potential is believed to be simple, needing primarily addition of glycosyltransferases to humanize the recombinant products. In this study, the native glycoproteome of Spodoptera frugiperda Sf9 and Trichoplusia ni High Five cells, examined using an LC-MS/MS approach, revealed not only which proteins are N-glycosylated, but indicated that the N-glycomes contain novel glucuronylated and phosphorylcholine-modified glycans, in addition to typical oligomannosidic and fucosylated structures. These data were corroborated by a parallel MALDI-TOF MS/MS analysis of N-glycosidase-released oligosaccharides. Molecular modelling analysis of one endogenous Sf9 glycoprotein correlated the occurrence of complex and oligomannosidic N-glycans with the accessibility of the occupied N-glycosylation sites. Further, we showed that the N-glycans of influenza haemagglutinins and SARS-CoV-2 spike glycoprotein produced in Spodoptera cells possess a number of glycan structures modified with phosphorylcholine, but core difucosylation was minimal; in contrast, the Trichoplusia-produced haemagglutinin had only traces of the former type, while the latter were dominant. Detection of phosphorylcholine on these glycoproteins correlated with binding to human C-reactive protein. In conclusion, not just oligomannosidic or truncated paucimannosidic N-glycans, but structures with immunogenic features occur on both natural and recombinant glycoproteins derived from insect cell lines.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100981"},"PeriodicalIF":6.1,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144023710","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}
James M Fulcher, Ashley N Ives, Shinya Tasaki, Shane S Kelly, Sarah M Williams, Thomas L Fillmore, Mowei Zhou, Ronald J Moore, Wei-Jun Qian, Ljiljana Paša-Tolić, Lei Yu, Shahram Oveisgharan, David A Bennett, Philip L De Jager, Vladislav A Petyuk
{"title":"Discovery of Proteoforms Associated with Alzheimer's Disease Through Quantitative Top-Down Proteomics.","authors":"James M Fulcher, Ashley N Ives, Shinya Tasaki, Shane S Kelly, Sarah M Williams, Thomas L Fillmore, Mowei Zhou, Ronald J Moore, Wei-Jun Qian, Ljiljana Paša-Tolić, Lei Yu, Shahram Oveisgharan, David A Bennett, Philip L De Jager, Vladislav A Petyuk","doi":"10.1016/j.mcpro.2025.100983","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100983","url":null,"abstract":"<p><p>The complex nature of Alzheimer's disease (AD) and its heterogenous clinical presentation has prompted numerous large-scale -omic analyses aimed at providing a global understanding of the pathophysiological processes involved. AD involves isoforms, proteolytic products, and post-translationally modified proteins such as amyloid beta (Aβ) and microtuble-associated protein tau. Top-down proteomics (TDP) directly measures these species, and thus, offers a comprehensive view of pathologically relevant proteoforms that are difficult to analyze using traditional proteomic techniques. Here, we broadly explored associations between proteoforms and clinicopathological traits of AD by deploying a quantitative TDP approach across frontal cortex of 103 subjects selected from the ROS and MAP cohorts. The approach identified 1,213 proteins and 11,782 proteoforms, of which 154 proteoforms had at least one significant association with a clinicopathological phenotype. One important finding included identifying Aβ C-terminal truncation state as the key property for differential association between amyloid plaques and cerebral amyloid angiopathy (CAA). Furthermore, various N-terminally truncated forms of Aβ had noticeably stronger association with amyloid plaques and global cognitive function. Additionally, we discovered six VGF neuropeptides that were positively associated with cognitive function independent of pathological burden. The database of brain cortex proteoforms provides a valuable context for functional characterization of the proteins involved in Alzheimer's disease and other late-onset brain pathologies.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100983"},"PeriodicalIF":6.1,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144027207","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}
Nazan Saner, Ceren Uzun, Büşra Aytül Akarlar, Sena Nur Özkan, Daniel Jon Geiszler, Ece Öztürk, Nurcan Tunçbağ, Nurhan Özlü
{"title":"Proximity labeling and SILAC based proteomic approach identifies proteins at the interface of homotypic and heterotypic cancer cell interactions.","authors":"Nazan Saner, Ceren Uzun, Büşra Aytül Akarlar, Sena Nur Özkan, Daniel Jon Geiszler, Ece Öztürk, Nurcan Tunçbağ, Nurhan Özlü","doi":"10.1016/j.mcpro.2025.100986","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100986","url":null,"abstract":"<p><p>Cell-cell interactions are critical for the growth of organisms and maintaining homeostasis. In the tumor microenvironment, these interactions promote cancer progression. Given their importance in healthy and diseased conditions, we have developed a method to analyze the cell-to-cell interactome. Our approach uses enzyme-catalyzed proximity labeling and SILAC-based proteomics to identify the proteins involved in cancer cell interactions. By targeting HRP to the outer leaflet of the plasma membrane in bait cells, we were able to label the neighboring prey cells and distinguish between the proteomes of bait and prey cells using SILAC labeling in a co-culture system. We mapped both the homotypic and heterotypic interactomes of epithelial and mesenchymal breast cancer cells. The enrichment of cell surface and extracellular proteins confirms the specificity of our methodology. We further verified selected hits from different cell-cell interactomes in co-cultures using microscopy. This method revealed prominent signaling pathways orchestrating homotypic and heterotypic interactions of epithelial and mesenchymal cells. It also highlights the importance of exosomes in these interactions. Our methodology can be applied to any type of cell-cell interaction in 2D co-culture or 3D tumor models.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100986"},"PeriodicalIF":6.1,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144012873","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}
Valdemaras Petrosius, Pedro Aragon-Fernandez, Tabiwang N Arrey, Jakob Woessman, Nil Üresin, Bauke de Boer, Jinyu Su, Benjamin Furtwängler, Hamish Stewart, Eduard Denisov, Johannes Petzoldt, Amelia C Peterson, Christian Hock, Eugen Damoc, Alexander Makarov, Vlad Zabrouskov, Bo T Porse, Erwin M Schoof
{"title":"Quantitative Label-Free Single-Cell Proteomics on the Orbitrap Astral MS.","authors":"Valdemaras Petrosius, Pedro Aragon-Fernandez, Tabiwang N Arrey, Jakob Woessman, Nil Üresin, Bauke de Boer, Jinyu Su, Benjamin Furtwängler, Hamish Stewart, Eduard Denisov, Johannes Petzoldt, Amelia C Peterson, Christian Hock, Eugen Damoc, Alexander Makarov, Vlad Zabrouskov, Bo T Porse, Erwin M Schoof","doi":"10.1016/j.mcpro.2025.100982","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100982","url":null,"abstract":"<p><p>Single-cell proteomics by mass spectrometry (scp-MS) holds the potential to provide unprecedented insights into molecular features directly linked to the cellular phenotype, while deconvoluting complex organisms into their basic building blocks. Tailored sample preparation that maximizes the extracted amount of material that is introduced into the mass spectrometer has rapidly propelled the field forward. However, the measured signal is still at the lower edge of detection approaching the sensitivity boundary of current instrumentation. Here, we investigate the capacity of the enhanced sensitivity of the Orbitrap Astral mass spectrometer to facilitate deeper proteome profiles from low-input to single-cell samples. We carry out a comprehensive data acquisition method survey to pinpoint which parameters provide most sensitivity. Furthermore, we explore the quantitative accuracy of the obtained measurements to ensure that the obtained abundances are in line with expected ground truth values. We culminate our technical exploration by generating small datasets from two cultured cell lines and a primary bone marrow sample, to showcase obtainable proteome coverage differences from different source materials. Finally, as a proof of concept we explore protein covariation to showcase how information on known protein complexes is captured inherently in our scp-MS data.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100982"},"PeriodicalIF":6.1,"publicationDate":"2025-05-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143990360","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}
Ashley M Frankenfield, Kevin L Yang, Wan Nur Atiqah Binti Mazli, Jamison Shih, Fengchao Yu, Edwin Lo, Alexey I Nesvizhskii, Ling Hao
{"title":"Benchmarking SILAC Proteomics Workflows and Data Analysis Platforms.","authors":"Ashley M Frankenfield, Kevin L Yang, Wan Nur Atiqah Binti Mazli, Jamison Shih, Fengchao Yu, Edwin Lo, Alexey I Nesvizhskii, Ling Hao","doi":"10.1016/j.mcpro.2025.100980","DOIUrl":"10.1016/j.mcpro.2025.100980","url":null,"abstract":"<p><p>Stable isotope labeling by amino acids in cell culture (SILAC) is a powerful metabolic labeling technique with broad applications and various study designs. SILAC proteomics relies on the accurate identification and quantification of all isotopic versions of proteins and peptides during both data acquisition and analysis. However, a comprehensive comparison and evaluation of SILAC data analysis platforms is currently lacking. To address this critical gap and offer practical guidelines for SILAC proteomics data analysis, we designed a comprehensive benchmarking pipeline to evaluate various in vitro SILAC workflows and commonly used data analysis software. Ten different SILAC data analysis workflows using five software packages (MaxQuant, Proteome Discoverer, FragPipe, DIA-NN, and Spectronaut) were evaluated for static and dynamic SILAC labeling with both DDA and DIA methods. For benchmarking, we used both in-house generated and repository SILAC proteomics datasets from HeLa and neuron culture samples. We assessed 12 performance metrics for SILAC proteomics including identification, quantification, accuracy, precision, reproducibility, filtering criteria, missing values, false discovery rate, protein half-life measurement, data completeness, unique software features, and speed of data analysis. Each method/software has its strengths and weaknesses when evaluated for these performance metrics. Most software reaches a dynamic range limit of 100-fold for accurate quantification of light/heavy ratios. We do not recommend using Proteome Discoverer for SILAC DDA analysis despite its wide use in label-free proteomics. To achieve greater confidence in SILAC quantification, researchers could use more than one software packages to analyze the same dataset for cross-validation. In summary, this study offers the first systematic evaluation of various SILAC data analysis platforms, providing practical guidelines to support decision-making in SILAC proteomics study design and data analysis.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100980"},"PeriodicalIF":6.1,"publicationDate":"2025-04-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144018351","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":"Uncommon N-Glycan Structures in Anhydrobiotic Tardigrades.","authors":"Hirokazu Yagi, Taiki Saito, Shih-Yun Guu, Nao Yamakawa, Shigeru Shimamura, Sachiko Kondo, Maho Yagi-Utsumi, Ken Takai, Jun-Ichi Furukawa, Yann Guerardel, Kay-Hooi Khoo, Kazuharu Arakawa, Koichi Kato","doi":"10.1016/j.mcpro.2025.100979","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100979","url":null,"abstract":"<p><p>We characterized the N-glycosylation profiles of anhydrobiotic tardigrades, R. varieornatus and H. exemplaris, identifying high-mannose, paucimannose, and complex-type oligosaccharides, while hybrid-type glycans were undetectable. Notably, paucimannose-type oligosaccharides accounted for 39% of the N-glycans in R. varieornatus and 17% in H. exemplaris, with a substantial proportion of them exhibiting fucosylation of the innermost GlcNAc via an α1,6-linkage. This core fucosylation pattern, common to all animals, was observed alongside a distinctive glycosylation signature prominently observed in tardigrades: complex-type glycans lacking galactosylation but containing α1,3-fucosylated GlcNAc at non-reducing termini. This structure was more prevalent in H. exemplaris, with 22 out of 87 identified glycoproteins expressing the Fucα1,3-GlcNAc motif, including eight induced during anhydrobiosis. Key glycoproteins such as Cu/Zn-superoxide dismutase and papilin, implicated in oxidative stress protection and extracellular matrix remodeling, were among those modified. Comparative analyses reveal that non-reducing terminal α1,3-fucosylation in tardigrades is distinct from the mammalian Lewis X antigen and similar structures found in invertebrates, suggesting a unique substrate specificity of fucosyltransferases in these species. Genomic analysis identified homologs of Fut9 and FucTC, indicating potential candidates responsible for this glycosylation pattern. Our findings provide new insights into the molecular mechanisms of glycosylation in tardigrades and its relevance to their extreme stress tolerance.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100979"},"PeriodicalIF":6.1,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144033567","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}