Molecular & Cellular Proteomics最新文献

{"title":"Data-independent Acquisition Proteomics Identifies Plasma Prostaglandin-H2D-isomerase as an Early Diagnostic Biomarker for STEMI and NSTEMI.","authors":"Hong-Mei Xue, Hai-Tao Hou, Yu Song, Huan-Xin Chen, Yun-Qiang Zhang, Wen-Tao Sun, Jie Zhou, Xiao-Lin Zhou, Na Sun, Qin Yang, Guo-Wei He","doi":"10.1016/j.mcpro.2025.100996","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100996","url":null,"abstract":"<p><p>Myocardial infarction (MI) including ST-elevated MI (STEMI) and non-ST-elevated MI (NSTEMI), remains a leading cause of death worldwide. This study aimed to identify the early diagnostic biomarkers for STEMI and NSTEMI. Plasma samples from 386 patients was classified into four groups: control (CON) (n=62), unstable angina (UA) (n=62), STEMI (n=182), and NSTEMI (n=80). The protein profiles were analyzed using data-independent acquisition (DIA)-based proteomics to identify differentially abundant proteins (DAPs) followed by bioinformatics analysis and ELISA validation. In STEMI, 93 DAPs were detected. Among the selected DAPs that were further validated in a new cohort of patients, prostaglandin-H2 D-isomerase (PTGDS) was elevated at the earliest onset time of STEMI (T1, 1.45h (95%CI: 1.16-1.73)) or NSTEMI (T1, 1.48h (95%CI: 0.97-1.98)) while the current biomarkers (hs-TnI, Myo, CKMB, and BNP) remained within normal ranges. The analysis of diagnostic indices for plasma PTGDS demonstrated a sensitivity of 63.95% and specificity of 65.38% in STEMI, 70% and 71.15% in NSTEMI. Moreover, AUC was 0.61 (95%CI: 0.53-0.69) in STEMI and 0.78 (95%CI: 0.70-0.86) in NSTEMI. The present study demonstrates that in MI patients, plasma PTGDS increases at an earlier stage of onset time than the current biomarkers with similar sensitivity and specificity. Therefore, PTGDS has high potential to be developed as an early diagnostic biomarker. In particular, PTGDS might be of greater clinical significance for patients suspected for NSTEMI, for which biomarker could be more effective in identifying high-risk patients suffering from MI at early stage.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100996"},"PeriodicalIF":6.1,"publicationDate":"2025-05-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144143004","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":"Data-driven extraction of human kinase-substrate relationships from omics datasets.","authors":"Benjamin Dominik Maier, Borgthor Petursson, Alessandro Lussana, Evangelia Petsalaki","doi":"10.1016/j.mcpro.2025.100994","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100994","url":null,"abstract":"<p><p>Phosphorylation forms an important part of the signalling system that cells use for decision making and regulation of processes such as cell division and differentiation. In human, >90% of identified phosphosites don't have annotations regarding the relevant upstream kinase. At the same time around 30% of kinases (as annotated in Uniprot) have no known target. This knowledge gap stresses the need to make large scale, data-driven computational predictions. In this study, we have created a machine learning-based model to derive a probabilistic kinase-substrate network from omics datasets. Our methodology displays improved performance compared to other state-of-the-art kinase-substrate prediction methods and provides predictions for more kinases. Importantly, it better captures new experimentally-identified kinase-substrate relationships. It can therefore allow the improved prioritisation of kinase-substrate pairs for illuminating the dark human cell signalling space. Our model is integrated into a web server, SELPHI_2.0, to allow unbiased analysis of phosphoproteomics data, facilitating the design of downstream experiments to uncover mechanisms of signal transduction across conditions and cellular contexts.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100994"},"PeriodicalIF":6.1,"publicationDate":"2025-05-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144093630","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":"Native Top-down analysis of membrane protein complexes directly from in vitro and native membranes.","authors":"Wonhyeuk Jung, Aniruddha Panda, Jaywon Lee, Snehasish Ghosh, Jared B Shaw, Kallol Gupta","doi":"10.1016/j.mcpro.2025.100993","DOIUrl":"https://doi.org/10.1016/j.mcpro.2025.100993","url":null,"abstract":"<p><p>Macromolecular organization of proteins and lipids in cellular membranes is fundamental to cell functionality. Recent advances in native mass spectrometry (nMS) have established it as a key analytical tool for capturing these associations. This typically necessitates the extraction of target membrane proteins from their physiological environments into detergent-like surroundings. In our recent studies using in vitro synthetic liposomes, we discovered that gas phase supercharging can selectively destabilize lipid bilayers and enable MS1 detection of embedded and associated protein-lipid complexes. Here, we further extend and apply this methodology to native cell-derived membrane vesicles. We demonstrate our ability to detect and ID protein complexes and their proteoforms directly from native membranes using supercharger-assisted pre-quadrupole activation followed by downstream native top-down MS/MS that combines both collision-based and electron capture-based fragmentations approaches. We first demonstrated this approach through native top-down identification of several integral membrane proteins from in vitro membranes. Subsequently, we developed a protocol to produce nMS-ready native membrane vesicles. Applying to E. coli total membranes, we generated nMS-ready vesicles and identified both integral and membrane-associated protein complexes of homomeric and heteromeric nature using our supercharging-enabled nTD-platform. For the hetero-pentameric BAM-complex, which includes the integral membrane protein BAM-A, we detected several lipidated proteoforms. For peripheral homo-dimeric DLDH, we identified bound endogenous metabolite co-factors. Furthermore, using BAM-complex, a crucial antibiotic target, we show how this platform could be utilized to study drug binding to membrane proteins directly from their native membranes.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100993"},"PeriodicalIF":6.1,"publicationDate":"2025-05-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144086652","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":"Proteomics and Phosphoproteomics Revealed Dysregulated Kinases and Potential Therapy for Liver Fibrosis.","authors":"Xinyu Cheng, Li Kang, Jinfang Liu, Qingye Wang, Zhenpeng Zhang, Li Zhang, Yuping Xie, Lei Chang, Daobing Zeng, Lantian Tian, Lingqiang Zhang, Ping Xu, Yanchang Li","doi":"10.1016/j.mcpro.2025.100991","DOIUrl":"10.1016/j.mcpro.2025.100991","url":null,"abstract":"<p><p>Liver fibrosis is the initial stage of most liver diseases, and it is also a pathological process involving the liver in the late stages of many metabolic diseases. Therefore, it is important to systematically understand the pathological mechanism of liver fibrosis and seek therapeutic approaches for intervention and treatment of liver fibrosis. Disordered proteins and their post-translational modifications, such as phosphorylation, play vital roles in the occurrence and development of liver fibrosis. However, the regulatory mechanisms that govern this process remain poorly understood. In this study, we analyzed and quantified the liver proteome and phosphoproteome of carbon tetrachloride-induced early liver fibrosis model in mice. Proteomic analysis revealed that the pathways involved in extracellular matrix recombination, collagen formation, metabolism and other related disorders, and protein phosphorylation modification pathways were also significantly enriched. In addition, Western blotting and phosphoproteomics demonstrated that phosphorylation levels were elevated in the context of liver fibrosis. A total of 13,152 phosphosites were identified, with 952 sites increased, whereas only 156 sites decreased. Furthermore, the upregulated phosphorylation sites, which exhibited no change at the proteome level, mainly shared a common [xxxSPxxx] motif. Consequently, the kinase-substrate analysis ascertained the overactive kinases of these upregulated substrates, which ultimately led to the identification of 13 significantly altered kinases within this dataset. These kinases were mainly cataloged into the STE, CMGC, and CAMK kinase families. Among them, STK4 (serine/threonine-protein kinase 4), GSK3α (glycogen synthase kinase 3α), and CDK11B (cyclin-dependent kinase 11B) were subsequently validated though cellular and animal experiments, and the results demonstrated that their inhibitors could effectively reduce the activation of hepatic stellate cells and extracellular matrix production. These kinases may represent potential therapeutic targets for liver fibrosis, and their inhibitors may serve as promising antihepatic fibrosis drugs.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100991"},"PeriodicalIF":6.1,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12181035/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144078703","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":"Chronic Alcohol Consumption Reprograms Hepatic Metabolism Through Organelle-Specific Acetylation in Mice.","authors":"Mirjavid Aghayev, Megan R McMullen, Serguei Ilchenko, Andrea Arias-Alvarado, Victor Lufi, Jack Mathis, Hannah Marchuk, Tsung-Heng Tsai, Guo-Fang Zhang, Laura E Nagy, Takhar Kasumov","doi":"10.1016/j.mcpro.2025.100990","DOIUrl":"10.1016/j.mcpro.2025.100990","url":null,"abstract":"<p><p>Posttranslational acetylation of proteins by acetyl-CoA is a crucial regulator of proteostasis and substrate metabolism. Ethanol metabolism in the liver induces protein accumulation, acetylation, and metabolic disruption. Although acetylation impacts enzyme activity and stability, its role in ethanol-related protein accumulation and metabolic dysfunction remains unclear. Using stable isotope-based proteomics, acetylomics, and metabolic profiling in a mouse model of chronic ethanol-induced liver injury, we demonstrate that ethanol induces hepatic steatosis, inflammation, oxidative stress, and proteinopathy linked to altered protein turnover. Ethanol increased the cytosolic protein turnover related to oxidative stress and detoxification, while reducing turnover of mitochondrial metabolic enzymes. It also elevated the acetylation of mitochondrial enzymes and nuclear histones with minimal cytosolic changes, impairing mitochondrial protein degradation. These changes were associated with altered levels of acyl-CoAs and acyl-carnitines, amino acids, and tricarboxylic acid cycle intermediates, reflecting impaired fatty acid oxidation, nitrogen disposal and tricarboxylic acid cycle activities. These results suggest that ethanol-induced acetylation contributes to liver injury and that targeting acetylation may offer treatment for alcohol-induced liver diseases.</p>","PeriodicalId":18712,"journal":{"name":"Molecular & Cellular Proteomics","volume":" ","pages":"100990"},"PeriodicalIF":6.1,"publicationDate":"2025-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144078222","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":"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":"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}

{"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":"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 through processed food or livestock 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 h 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 the 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}

{"title":"Laser-Induced Rehydration of Cryo-Landed Proteins Restores Native Structure.","authors":"Keaton L Mertz, Drew 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":"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 long-standing 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. Furthermore, 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":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12171540/pdf/","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":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"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":"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 also 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 modeling 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 hemagglutinins 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 hemagglutinin had only traces of the former type, while the latter was 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":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12166434/pdf/","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":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}