Proteomes

30 pages, 3069 KB

Open AccessReview

From Proteomic Signatures to Candidate Endotypes in Obesity, Type 2 Diabetes, MASLD/MASH, and MetALD: Study Designs, Bioinformatics, and Biostatistical Strategies

by Zhennan Wu, Sachin Anil Ghag, Md Hasan Imam Shihab, Aatman Pushkarkumar Vasoya, Vinamrata Sharma, Jacob Patton Hickman, Yijie Wang, Xiaoqing Huang and Menghao Huang

Proteomes 2026, 14(3), 49; https://doi.org/10.3390/proteomes14030049 (registering DOI) - 20 Sep 2026

Abstract

Obesity, type 2 diabetes (T2D), metabolic dysfunction-associated steatotic liver disease (MASLD), metabolic dysfunction-associated steatohepatitis (MASH), and dual-etiology metabolic dysfunction-associated alcohol-related liver disease (MetALD) form an overlapping metabolic dysfunction spectrum, rather than a single linear disease sequence. Proteomics offers a functional readout of this [...] Read more.

Obesity, type 2 diabetes (T2D), metabolic dysfunction-associated steatotic liver disease (MASLD), metabolic dysfunction-associated steatohepatitis (MASH), and dual-etiology metabolic dysfunction-associated alcohol-related liver disease (MetALD) form an overlapping metabolic dysfunction spectrum, rather than a single linear disease sequence. Proteomics offers a functional readout of this spectrum by measuring proteins, proteoforms, and protein species involved in tissue injury, inflammation, metabolic stress, and inter-organ communication. This review asks how proteomic data can support mechanism-based stratification, rather than simply generate disease-associated signatures. We summarize advances in circulating and tissue-based proteomics across obesity, T2D, MASLD/MASH, and MetALD, highlighting shared and disease-specific pathways such as mitochondrial dysfunction, extracellular matrix remodeling, immune activation, proteostasis stress, and endocrine crosstalk. We emphasize that proteomic clusters should be considered candidate endotypes only when they are reproducible, mechanistically coherent, linked to tissue or causal evidence, and clinically informative. We also evaluate bioinformatics and biostatistical strategies needed for reliable interpretation, including preprocessing, missing-data handling, normalization, longitudinal modeling, multi-omics integration, protein quantitative trait locus (pQTL) analysis, colocalization, and Mendelian randomization. Finally, we discuss how proteoforms, post-translational modifications (PTMs), and platform-dependent proteome complexity shape interpretation. Together, these concepts provide practical guidance for moving from proteomic signatures to candidate endotypes and for prioritizing clinically useful biomarkers and therapeutic targets. Full article

(This article belongs to the Section Proteomics of Human Diseases and Their Treatments)

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19 pages, 1768 KB

Open AccessReview

Biological Functions and Applications of Exosomes from Periodontal Ligament Stem Cells

by Omer Tarik Ozdemir, Hideki Sugii, Bara Mardini and Hidefumi Maeda

Proteomes 2026, 14(3), 48; https://doi.org/10.3390/proteomes14030048 - 10 Sep 2026

Viewed by 242

Abstract

Periodontal regeneration requires the reconstruction of root cementum, periodontal ligament, and alveolar bone, which are frequently disrupted by periodontitis, trauma, and other inflammatory conditions. Although periodontal ligament stem cells (PDLSCs) have been considered promising for periodontal tissue engineering because of their multipotency, self-renewal [...] Read more.

Periodontal regeneration requires the reconstruction of root cementum, periodontal ligament, and alveolar bone, which are frequently disrupted by periodontitis, trauma, and other inflammatory conditions. Although periodontal ligament stem cells (PDLSCs) have been considered promising for periodontal tissue engineering because of their multipotency, self-renewal capacity, and immunomodulatory activity, direct cell-based therapy still faces various limitations, including donor variability, culture-related changes, delivery difficulties, and safety concerns. Exosomes and small extracellular vesicles derived from PDLSCs have therefore attracted attention as potential cell-free mediators of PDLSC paracrine activity. PDLSC-derived exosomes carry bioactive molecules, including proteins, lipids, messenger RNAs, and microRNAs, and can influence recipient cell behavior in periodontal and bone regenerative environments. In vitro and preclinical animal studies indicate that PDLSC-derived exosomes are involved in osteogenesis, periodontal attachment repair, cementogenesis, angiogenesis, mechanotransduction, cell proliferation/survival, and immune regulation. Their biological effects are closely related to the condition of their parental PDLSCs, their cargo, and the exosome isolation and characterization methods used. In this review, we summarize recent findings on the isolation strategies used, biological functions of, and therapeutic potential of PDLSC-derived exosomes, with particular focus on protein cargo, proteomic signatures, proteoform-related interpretation, and microRNA-mediated regulation. We also discuss current challenges in exosome isolation, characterization, delivery, stability, and reporting standards for future periodontal regenerative applications. The findings discussed throughout are preclinical, and their translation to clinical settings remains to be explored. Full article

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21 pages, 2149 KB

Open AccessArticle

A Circulating Proteomic Signature of Allostatic Load Is Associated with Multisystem Disease and Mortality

by Yufan Guan, Jie Shen, Li Li, Kai Zhang, Song Liu, Qianqian Zhu, Philip I. Chow, Hongmei Jiang, Jing Li, Lifang Hou and Hua Zhao

Proteomes 2026, 14(3), 47; https://doi.org/10.3390/proteomes14030047 - 10 Sep 2026

Viewed by 189

Abstract

Background: Chronic stress contributes to the development of cardiometabolic, malignant, and other chronic diseases through cumulative multisystem physiological dysregulation, conceptualized as allostatic load (AL). However, traditional AL relies on heterogeneous clinical biomarkers that are frequently unavailable or inconsistently collected in large cohorts, limiting [...] Read more.

Background: Chronic stress contributes to the development of cardiometabolic, malignant, and other chronic diseases through cumulative multisystem physiological dysregulation, conceptualized as allostatic load (AL). However, traditional AL relies on heterogeneous clinical biomarkers that are frequently unavailable or inconsistently collected in large cohorts, limiting reproducibility and translational utility. We developed and validated ProAL50, a proteomics-based measure of AL derived from 50 circulating proteins. Methods: Using high-dimensional plasma proteomic data from the UK Biobank, we constructed ProAL50 via penalized regression and stability selection and externally validated its construct in the Coronary Artery Risk Development in Young Adults (CARDIA) Study. Results: Among 37,089 UK Biobank participants, ProAL50 correlated strongly with traditional AL in the held-out test set (r = 0.72) and showed moderate external construct validity in CARDIA (r = 0.62). ProAL50 closely mirrored traditional AL in its associations with sociodemographic characteristics, lifestyle behaviors, physical and mental health, inflammation, and biological aging, supporting construct validity. In prospective analyses, ProAL50 showed associations with incident chronic disease, including type 2 diabetes, ischemic heart disease, chronic lung disease, chronic liver disease, chronic kidney disease, overall cancer, and all-cause and cause-specific mortality that were broadly comparable to traditional AL. Representative associations included incident type 2 diabetes (HR per SD = 2.72, 95% CI 2.61–2.84) and chronic liver disease (HR = 2.12, 95% CI 1.96–2.29). Conclusions: These findings support ProAL50 as a biologically informative, protein-based surrogate of AL for research use that complements traditional AL, pending further external validation and assay standardization. Full article

(This article belongs to the Section Proteomics of Human Diseases and Their Treatments)

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10 pages, 3512 KB

Open AccessArticle

The “2DE-Pattern” Database for Inventory of Proteoform Profiles: 2026 Upgrade and Update on Outcomes

by Stanislav Naryzhny, Nikolay Klopov, Natalia Ronzhina, Elena Zorina and Olga Legina

Proteomes 2026, 14(3), 46; https://doi.org/10.3390/proteomes14030046 - 7 Sep 2026

Viewed by 286

Abstract

Background: Modern proteomics faces a critical bottleneck: the vast discrepancy between the number of genes in the human genome and the exponentially greater variety of functional proteoforms that actually drive biological processes. Methods: Our paper addresses the urgent need for high-resolution systematic mapping [...] Read more.

Background: Modern proteomics faces a critical bottleneck: the vast discrepancy between the number of genes in the human genome and the exponentially greater variety of functional proteoforms that actually drive biological processes. Methods: Our paper addresses the urgent need for high-resolution systematic mapping of these proteoforms, arguing that the true frontier of molecular biology lies in the precise identification and categorization of protein variants. It centers on the development and expansion of the “2DE-pattern” database, a specialized platform designed to bridge the gap between theoretical protein sequences and the physical reality of proteins as captured through two-dimensional electrophoresis (2DE). The “2DE-pattern” database is based on information obtained by separation of proteoforms using 2DE followed by shotgun ESI LC-MS/MS. It was launched in 2020, contains multiple isoform-centric patterns of proteoforms, and can be freely used. Results: Here, we report the additional data and all updates that were added into this database. Also, the database was upgraded to be more research-oriented. Tools were incorporated into the database to allow convenient comparative analysis of the data. Conclusions: New additions and enhancements now allow us to consider our database a knowledge base. Full article

(This article belongs to the Section Proteoform Analysis (Top-Down and Bottom-Up))

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28 pages, 19515 KB

Open AccessArticle

Integrative Proteomic Analysis Implicates Inhibition of Intracellular Protein Trafficking in Therapy-Induced Migrastasis in Prostate Cancer

by Weining Chen, Saadyeh Rashidi, Henry C.-H. Law, Fangfang Qiao, Johnny W. Zigmond, Katelyn L. O’Neill, Nicholas T. Woods, Chittibabu Guda and Raymond C. Bergan

Proteomes 2026, 14(3), 45; https://doi.org/10.3390/proteomes14030045 - 28 Aug 2026

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Abstract

Background: Dysregulated cell migration leading to metastasis remains the primary cause of cancer-related mortality. It has been challenging to understand how cells regulate migration. We have previously created the first selective inhibitor of cell migration, KBU2046. Here, we use it as a probe [...] Read more.

Background: Dysregulated cell migration leading to metastasis remains the primary cause of cancer-related mortality. It has been challenging to understand how cells regulate migration. We have previously created the first selective inhibitor of cell migration, KBU2046. Here, we use it as a probe to identify regulatory processes. Methods: Metastatic and primary human prostate cancer cells were treated for different times and at different concentrations with KBU2046. Immunofluorescent microscopy examined protein localization in cells. Label-free mass spectrometry (MS) was performed on total cell proteins, Tandem Mass Tag (TMT) labeling MS was used on membrane fractions, and temporal phosphoproteomic profiling was performed. Results were analyzed with a suite of bioinformatic tools. Results: KBU2046-induced migrastasis is associated with the accumulation of activated integrin β1 into focal adhesions. Whole-cell proteomics demonstrated suppression of processes that mediate intracellular protein trafficking and increases in mitochondrial energy-generation signatures. Evaluation of the membrane fraction identified increases in membrane repair and maintenance processes and decreases in those that drive motility. Temporal- and concentration-dependent phosphoproteomic profiling revealed that KBU2046 initiates a dynamic, cascading sequence of transient signaling waves rather than a static block. Conclusions: KBU2046-induced migrastasis appears to operate through spatial decoupling rather than structural degradation. By restricting the intracellular trafficking machinery required for receptor recycling, KBU2046 limits focal adhesion turnover, providing, in PC3 prostate cancer cells, a correlative framework to inhibit metastatic dissemination independent of direct cytotoxicity. Full article

(This article belongs to the Section Proteomics of Human Diseases and Their Treatments)

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42 pages, 1321 KB

Open AccessReview

Beyond Protein Abundance: Proteoform Diversity and Phosphoproteome Remodeling in Maize (Zea mays L.) Responding to Drought and Heat Stresses

by Jan Bocianowski

Proteomes 2026, 14(3), 44; https://doi.org/10.3390/proteomes14030044 - 27 Aug 2026

Viewed by 266

Abstract

Drought and heat stresses are among the most important environmental constraints limiting maize (Zea mays L.) productivity worldwide. Over the past two decades, advances in mass spectrometry-based proteomics have generated extensive datasets describing proteins with altered abundance in maize tissues exposed to [...] Read more.

Drought and heat stresses are among the most important environmental constraints limiting maize (Zea mays L.) productivity worldwide. Over the past two decades, advances in mass spectrometry-based proteomics have generated extensive datasets describing proteins with altered abundance in maize tissues exposed to water deficit and elevated temperatures. These studies have identified numerous stress-responsive proteins involved in photosynthesis, energy metabolism, antioxidant defense, proteostasis, signaling pathways, and cellular homeostasis. In parallel, phosphoproteomic investigations have revealed extensive remodeling of phosphorylation networks that regulate stress perception, signal transduction, and adaptive responses. However, most available studies remain focused on protein-level observations and provide limited insight into the molecular diversity underlying stress adaptation. This review synthesizes current knowledge on the proteomic and phosphoproteomic responding of maize to drought and heat stresses through the emerging perspective of proteoform biology. We discuss how phosphorylation, oxidative modifications, proteolytic processing, alternative splicing, and genetic variation contribute to proteoform generation and expand the functional complexity of the maize stress proteome. Particular emphasis is placed on the integration of quantitative proteomics, phosphoproteomics, and proteogenomics as complementary approaches for characterizing stress-responsive molecular networks. We further evaluate current methodological limitations, including the predominance of bottom-up workflows, the underrepresentation of combined-stress studies and reproductive tissues, and the limited application of proteoform-resolved analytical strategies. We propose that future advances in maize stress biology will require a transition from protein-centered analyses toward proteoform-centered investigations capable of resolving functionally distinct molecular species. The integration of top-down proteomics, phosphoproteomics, proteogenomics, and systems-level approaches is expected to provide a more comprehensive understanding of stress adaptation mechanisms and facilitate the identification of molecular determinants of climate resilience. Such efforts may ultimately support the development of maize cultivars better adapted to increasingly challenging environmental conditions. Full article

(This article belongs to the Special Issue Plant Genomics and Proteomics)

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20 pages, 1984 KB

Open AccessArticle

Urinary Proteome Profiling by Several Methods Identifies Titin as the Most-Differentiating Noninvasive Urinary Biomarker of Disease Severity in Becker Muscular Dystrophy

by Kimchi K. Le, Emily H. Canessa, Corrine M. Stahura, Jenna Mayers, Katie A. Edwards, Marissa Barbieri, Eric P. Hoffman and Yetrib Hathout

Proteomes 2026, 14(3), 43; https://doi.org/10.3390/proteomes14030043 - 25 Aug 2026

Viewed by 549

Abstract

Background: Beckers muscular dystrophy (BMD) is a clinically heterogeneous dystrophinopathy caused by in-frame mutations in the dystrophin gene, resulting in variable disease severity. This variability limits the effectiveness of standardized functional measures for tracking progression. Circulating biomarkers, including urinary titin fragments generated during [...] Read more.

Background: Beckers muscular dystrophy (BMD) is a clinically heterogeneous dystrophinopathy caused by in-frame mutations in the dystrophin gene, resulting in variable disease severity. This variability limits the effectiveness of standardized functional measures for tracking progression. Circulating biomarkers, including urinary titin fragments generated during muscle injury, offer a promising non-invasive approach for assessing disease status. Methods: Mass spectrometry-based urinary proteomic profiling identified titin fragments as candidate biomarkers of muscle injury in BMD. These titin fragments were validated by two independent methods, targeted mass spectrometry and ELISA, using subsets of ambulatory BMD, non-ambulatory BMD and age matched healthy volunteers. Results: Urinary titin levels were significantly elevated by 4.56-fold and 2.32-fold in ambulatory and non-ambulatory BMD patients, respectively, compared with healthy controls. Titin levels also distinguished ambulatory from non-ambulatory BMD patients, demonstrating potential utility for monitoring disease progression and therapeutic response. Conclusions: Urinary titin fragments represent a promising non-invasive biomarker for BMD, with potential applicability to related neuromuscular disorders and clinical trials evaluating disease progression and treatment efficacy. Full article

(This article belongs to the Special Issue Clinical Proteomics: Unravelling Drug and Disease Mechanisms (Fifth Edition))

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31 pages, 12452 KB

Open AccessArticle

DOG1-Mediated Priming Followed by Environmentally Tunable Plasticity: A Two-Phase Model for Dormancy Establishment in Xanthium strumarium

by Iman Nemati, Somayeh Gholizadeh, Dinakaran Elango, Sara Hamzelou, Karthik Shantharam Kamath, Mohammad Sedghi, Reza Tavakkol Afshari and Paul A. Haynes

Proteomes 2026, 14(3), 42; https://doi.org/10.3390/proteomes14030042 - 21 Aug 2026

Viewed by 315

Abstract

Background: Seed dormancy is crucial for plant survival and agricultural productivity, yet its molecular mechanisms, particularly the role of maternal effects, remain poorly understood. Methods: In this study, we applied a SWATH-based, label-free, quantitative shotgun proteomic mass spectrometry approach to investigate the temporal [...] Read more.

Background: Seed dormancy is crucial for plant survival and agricultural productivity, yet its molecular mechanisms, particularly the role of maternal effects, remain poorly understood. Methods: In this study, we applied a SWATH-based, label-free, quantitative shotgun proteomic mass spectrometry approach to investigate the temporal dynamics of dormancy establishment in Xanthium strumarium, a wild plant with two seeds in one burr that, despite sharing the same genetic and environmental conditions, exhibit distinct dormancy states. Results: Our data show that dormant seeds undergo coordinated metabolic suppression, marked by a decrease in energy metabolism, cell cycle arrest, and auxin signaling, explaining their smaller size. Simultaneously, dormant seeds exhibit metabolic re-prioritization towards fatty acid desaturation, cell wall modification, and an active epigenetic program stabilized by dormancy-promoting factors alongside a transcriptionally quiescent state in early–mid development. However, in the late developmental stage, molecular signaling pathways showed a recalibration distinguished by changes in seed metabolism (such as carbon–nitrogen reallocation, sulfur assimilation, and GABA production), hormonal fluctuations, and epigenetic regulation. Notably, previously reported high DOG1 transcript abundance, together with the absence of detectable DOG1 protein in the proteomic dataset, suggests that post-transcriptional mechanisms may contribute to DOG1 regulation. Conclusions: Based on these findings and the available literature, we propose a framework whereby dormancy establishment occurs in two phases: an early DOG1-mediated priming phase followed by a temperature-sensitive plasticity phase during seed maturation. Full article

(This article belongs to the Special Issue Plant Genomics and Proteomics)

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42 pages, 11117 KB

Open AccessArticle

A Propeller with a Flexible Twist: A Computational Analysis of Intrinsically Disordered Regions in PIEZO Gating and PIEZO-Associated Channelopathies

by Shivam Shukla, Mason Elzy, Abiral Shrestha and Vladimir N. Uversky

Proteomes 2026, 14(3), 41; https://doi.org/10.3390/proteomes14030041 - 11 Aug 2026

Viewed by 652

Abstract

Background: Mechanosensitive ion channels PIEZO1 and PIEZO2 are key mediators of mechanotransduction, which converts physical forces into cellular signals involved in proprioception, touch, vascular function, and other physiological processes. Mutations in human PIEZO proteins are linked to various diseases, such as hereditary xerocytosis, [...] Read more.

Background: Mechanosensitive ion channels PIEZO1 and PIEZO2 are key mediators of mechanotransduction, which converts physical forces into cellular signals involved in proprioception, touch, vascular function, and other physiological processes. Mutations in human PIEZO proteins are linked to various diseases, such as hereditary xerocytosis, lymphatic dysplasia, and proprioceptive dysfunction. However, the role of intrinsic disorder in the regulation of these proteins and their susceptibility for disease-associated mutations remains unclear. Methods: We analyzed canonical human PIEZO1 and PIEZO2 protein sequences using machine learning, neural network, and energy-based disorder predictors, together with the prediction of disorder-mediated binding regions, phase separation propensity, interaction networks, evolutionary conservation, clinically annotated human variants, and peptide structural modeling. Results: Both proteins showed moderate intrinsic disorder, with PIEZO2 having slightly greater disorder propensity and higher predicted phase separation potential. Intrinsically disordered regions frequently overlapped binding-prone segments and post-translational modification sites, supporting regulatory functions. Evolutionary comparisons showed strong conservation of PIEZO proteins, while selected disordered regions retained disorder propensity despite greater sequence variability. Disease-causing variants mainly affected the ordered regions of both proteins, whereas disordered regions contained proportionally more benign variants and relatively few pathogenic mutations. The modeling of mutations within disordered hotspots showed altered local conformational tendencies, indicating that some disease variants may disrupt dynamic interaction interfaces rather than global structure. Interaction network analysis linked both proteins to enriched mechanotransduction, ion transport, and cytoskeletal pathways. Conclusions: Overall, our findings identify intrinsic disorder as an underappreciated feature of PIEZO channel biology and provide a framework for interpreting PIEZO-associated channelopathies. PIEZO proteins also perfectly illustrate the proteoform concept, where one gene yields a highly diverse kit of mechanosensitive molecular tools. While humans only have two primary PIEZO genes (PIEZO1 and PIEZO2), the body generates a vast array of functional variations. Full article

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19 pages, 931 KB

Open AccessArticle

Comparative Analysis of Progesterone Secretion and Plasma Proteome Across the Pregnant and Non-Pregnant Luteal Phase of the Koala (Phascolarctos cinereus)

by Brooke E. Hartley, Stephen D. Johnston, Yolande Campbell, Kerry Fanson, Vere Nicolson, Ashleigh Neal and Taylor Pini

Proteomes 2026, 14(3), 40; https://doi.org/10.3390/proteomes14030040 - 6 Aug 2026

Viewed by 718

Abstract

Background: Koalas are a vulnerable marsupial species with unique reproductive traits. Efforts to develop assisted breeding technologies have been hindered by a limited understanding of maternal recognition of pregnancy and physiological changes induced by the foeto-placental unit. Differences in the reproductive physiology of [...] Read more.

Background: Koalas are a vulnerable marsupial species with unique reproductive traits. Efforts to develop assisted breeding technologies have been hindered by a limited understanding of maternal recognition of pregnancy and physiological changes induced by the foeto-placental unit. Differences in the reproductive physiology of pregnant and non-pregnant koalas were examined to investigate the possibility of maternal recognition and identify potential pregnancy and/or embryonic loss biomarkers. Methods: Koalas were separated into three groups: pregnant (n = 4 cycles from three females), mated but non-parturient (n = 4 cycles from three females), and gonadotropin-releasing hormone (GnRH) agonist-treated females (n = 7). Plasma was collected on day of mating/GnRH injection (D0) and on multiple subsequent days. Progesterone concentrations were measured by enzyme immunoassay, and plasma proteomes were analysed using filter-aided sample preparation followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), employing sequential window acquisition of all theoretical fragment ion spectra. Results: Ovulation induction mechanisms influenced peri-ovulatory progesterone secretion (pregnant 40.7 ± 3.3 ng/mL vs. GnRH-treated 15.7 ± 1.8 ng/mL), with no significant differences in progesterone that occurred later in the luteal phase. LC-MS/MS identified 158 proteins, representing the first koala plasma proteome. Leucine-rich alpha-2-glycoprotein (LRG1) was significantly elevated at D2 in pregnant females compared to GnRH-treated females, and pregnant D9 and D19. In pregnant females, fibronectin (FN1) was significantly more abundant at D19 compared to D9 but not significantly different between treatments. Conclusions: These preliminary findings provide foundational data for further investigation into maternal recognition and pregnancy/embryonic loss in koalas. Full article

(This article belongs to the Section Animal Proteomics)

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22 pages, 6583 KB

Open AccessArticle

Proteomic Analysis of Paraffin-Embedded Intestines from Schistosoma mansoni Infection in Mice: Highlighting Molecular Players During Acute Schistosomiasis

by Lara Geralda Magela dos Santos Vieira, Ana Flávia Pinho Souza, Camilo Elber Vital, Dávila Regina Pacheco Silva, Flávia de Souza Marques, Gustavo Gonçalves Silva, Paula Melo de Abreu Vieira, R Alan Wilson and William Castro-Borges

Proteomes 2026, 14(3), 39; https://doi.org/10.3390/proteomes14030039 - 29 Jul 2026

Viewed by 1326

Abstract

Background: Adult Schistosoma mansoni parasites inhabit the hepatic portal system of the vertebrate host, their deposited eggs causing granulomatous pathology in both the intestines and liver. In the intestines, egg secretions drive inflammatory processes involved in extravasation to the gut lumen. Methods: Here, [...] Read more.

Background: Adult Schistosoma mansoni parasites inhabit the hepatic portal system of the vertebrate host, their deposited eggs causing granulomatous pathology in both the intestines and liver. In the intestines, egg secretions drive inflammatory processes involved in extravasation to the gut lumen. Methods: Here, we investigate parasite-host interactions in a mouse model during the acute phase of a patent infection at 5 and 7 weeks, using parallel histological and proteomic analysis of paraffin-embedded ileal tissues. Results: Histology at week 7 showed infected animals had more inflammation and longer villi than at week 5, as well as more Goblet cells reflecting enhanced mucus production. LC-MS/MS analysis of deparaffinized ileal sections, subjected to in-solution tryptic digestion, revealed a total of 1615 protein groups. Differentially abundant proteins were found early at week 5, coinciding with the onset of egg deposition. A contrasting scenario, dominated by upregulation of protein components from the innate and adaptive immune systems, was seen at week 7; at this point, egg migration and excretion are underway. Among the proteins were mast cell proteases, fibrinogens, arginase-1, and molecules associated with extracellular matrix remodeling. Conclusions: Our findings reflect intestinal proteome changes likely participating in S. mansoni egg passage from the vascular bed to the intestinal lumen. Full article

(This article belongs to the Section Animal Proteomics)

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27 pages, 1926 KB

Open AccessArticle

Proteomic Mediators Linking Autoimmune Diseases to Major Adverse Cardiovascular Events: Insights from the UK Biobank

by Jingwen Huang, Chang Liu, Laurence S. Sperling, Arshed A. Quyyumi and Yan V. Sun

Proteomes 2026, 14(3), 38; https://doi.org/10.3390/proteomes14030038 - 24 Jul 2026

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Abstract

Background: Autoimmune diseases (AIDs) are associated with increased cardiovascular risk. However, specific protein mediators linking AIDs to major adverse cardiovascular events (MACE) and cardiovascular death (CV death) remain unexplored. This study identifies proteomic mediators linking AIDs to MACE via high-dimensional mediation analysis in [...] Read more.

Background: Autoimmune diseases (AIDs) are associated with increased cardiovascular risk. However, specific protein mediators linking AIDs to major adverse cardiovascular events (MACE) and cardiovascular death (CV death) remain unexplored. This study identifies proteomic mediators linking AIDs to MACE via high-dimensional mediation analysis in the UK Biobank. Methods: We used UK Biobank data with proteomic profiling by Olink platform. Participants with prevalent myocardial infarction (MI), stroke, and heart failure at baseline were excluded. AIDs were categorized into musculoskeletal (MSK), vasculitis, gastrointestinal (GI), neurologic, and rheumatic fever subsets. Fine–Gray models assessed associations between AIDs and MACE and CV death. Proteome-wide association studies identified proteins associated with both AIDs and cardiovascular outcomes. High-dimensional mediation analysis (HIMA) explored protein-mediated pathways. All models adjusted for age, sex, lipids, BMI, smoking, hypertension, diabetes, chronic kidney disease, atrial fibrillation, and coronary artery disease. Results: Among 400,633 participants (median follow-up 14.5 years, 44.8% male), AIDs were present in 28,754 (7.2%). All AID categories were associated with increased MACE (sHR: MSK 1.34, vasculitis 1.67, GI 1.20, neurologic 1.33, rheumatic fever 1.38; all p < 0.001). For CV death, MSK, vasculitis, and rheumatic fever showed increased risk (sHR 1.34, 1.78, 1.51; all p ≤ 0.004), but not GI or neurologic AIDs. In 43,599 participants with proteomic data, HIMA identified 66 and 32 unique potential mediators linking AIDs to MACE and CV death, respectively. Four proteins (Growth Differentiation Factor 15, Interleukin-15, urokinase plasminogen activator receptor, and Tenascin C) mediated the AID-MACE relationship across multiple AID categories. Growth Differentiation Factor 15 and Interleukin-15 were shared mediators for CV death. Conclusions: This proteomic analysis identifies specific proteins that may mediate the association between AIDs and adverse cardiovascular outcomes, offering mechanistic insights into immune-related cardiovascular risk. These findings are hypothesis-generating and require replication and validation before the identified proteins can be considered causal mediators or adopted for clinical risk stratification. Full article

(This article belongs to the Section Proteomics of Human Diseases and Their Treatments)

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27 pages, 10788 KB

Open AccessArticle

Proteomic Profiling Reveals Region-Specific Brain Responses During Acclimation to Elevated Water Temperature in Atlantic Salmon (Salmo salar L.)

by Manojkumar Chandraprakasham, Gianluca Amoroso, Chris G. Carter, Chloe J. English, Lambertus Koster, Richard Wilson, Richard S. Taylor and Omar Mendoza-Porras

Proteomes 2026, 14(3), 37; https://doi.org/10.3390/proteomes14030037 - 23 Jul 2026

Cited by 1 | Viewed by 841

Abstract

Background: Increasing summer seawater temperatures pose challenges for Atlantic salmon aquaculture, while brain region-specific responses to elevated temperature remain poorly understood. Methods: Atlantic salmon in the warm treatment (WT) underwent thermal ramping from 15 °C to 19 °C, with mortality observed at the [...] Read more.

Background: Increasing summer seawater temperatures pose challenges for Atlantic salmon aquaculture, while brain region-specific responses to elevated temperature remain poorly understood. Methods: Atlantic salmon in the warm treatment (WT) underwent thermal ramping from 15 °C to 19 °C, with mortality observed at the end of week 4 (15.78%). The WT temperature was subsequently reduced to 18 °C in week 5, while the control treatment (CT) was adjusted from 15 °C to 14 °C, and the WT (18 °C) and CT (14 °C) conditions were maintained thereafter. At the end of the trial (week 10), six brain regions, including the cerebellum (CBE), hypothalamus (HYP), medulla oblongata (MED), optic tectum (OPT), pituitary gland (PIT), and telencephalon (TEL), were analysed using data-independent acquisition mass spectrometry-based proteomics. Results: Over 9000 protein groups were identified per brain region, and exploratory differential abundance analysis revealed predominantly region-specific protein abundance changes. Increased SERPINH1 (HSP47) abundance was observed in HYP, PIT, and TEL, suggesting roles in protein-folding and stress regulation. Functional enrichment analyses indicated differential regulation of translation, transcription, energy metabolism, and metabolic pathways across brain regions in the WT group. Conclusions: This study provides a brain region-specific proteomic resource for Atlantic salmon and advances understanding of molecular responses associated with recovery from a temperature reduction (19 °C to 18 °C) and subsequent thermal adjustment at 18 °C. Full article

(This article belongs to the Section Animal Proteomics)

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18 pages, 7604 KB

Open AccessArticle

Protein Language Model Embeddings Reveal Proteome-Scale Ortholog Divergence Relevant to Cross-Species Pharmacology

by Taichi Endoh, Gerry Amor Camer, Kotetsu Kayama, Daiji Endoh and Hiroki Teraoka

Proteomes 2026, 14(3), 36; https://doi.org/10.3390/proteomes14030036 - 23 Jul 2026

Viewed by 925

Abstract

Background: Comparative proteome analysis can reveal functional conservation and divergence among orthologous proteins, with important implications for pharmacology and toxicology. Protein language models (PLMs) may capture sequence-derived functional relationships beyond what conventional alignment metrics capture. Methods: Orthologous proteins from Danio rerio and Danio [...] Read more.

Background: Comparative proteome analysis can reveal functional conservation and divergence among orthologous proteins, with important implications for pharmacology and toxicology. Protein language models (PLMs) may capture sequence-derived functional relationships beyond what conventional alignment metrics capture. Methods: Orthologous proteins from Danio rerio and Danio aesculapii were compared using embeddings generated by the Evolutionary Scale Modeling 2 (ESM-2) protein language model. Reciprocal best-hit inference identified 68,971 high-confidence ortholog pairs, of which 51,086 were available for embedding-based analysis. PLM divergence was quantified using cosine distance and evaluated using length-matched and bitscore-matched random controls, Gene Ontology graph-distance analysis, and localized domain-level comparisons. Results: Ortholog pairs showed strong global conservation, with a median PLM distance of 0.000487, whereas randomized controls exhibited substantially greater divergence. Increasing Gene Ontology graph distance broadened PLM-distance distributions, and leaf–parent comparisons demonstrated significant functional ordering (Wilcoxon p = 2.44 × 10⁻⁴). Local analyses revealed increased divergence in pathophysiologically relevant regions of aryl hydrocarbon receptor (AHR) and potassium channel proteins. Conclusions: PLM embeddings provide a scalable framework for comparative proteome characterization, complement conventional sequence-based analyses, and prioritize orthologs or protein regions with elevated functional divergence for experimental validation in cross-species pharmacology, toxicology, and systems biology. Full article

(This article belongs to the Section Proteome Bioinformatics)

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18 pages, 1943 KB

Open AccessArticle

Lyophilization Prior to Homogenisation and Extraction Increases Membrane Protein Detection in Gram-Negative Bacterial Proteomic Analyses

by Breyer Woodland, Luke A. Farrell, Matthew B. O’Rourke and Matthew P. Padula

Proteomes 2026, 14(3), 35; https://doi.org/10.3390/proteomes14030035 - 15 Jul 2026

Viewed by 677

Abstract

Background: Multi-drug resistant Gram-negative bacteria (GNB) are major contributors to the antimicrobial resistance (AMR) burden. AMR mechanisms are primarily mediated by proteoforms; therefore, proteomic analyses of GNB offers a significant advantage in understanding the mechanisms of AMR. A large portion of these mechanisms [...] Read more.

Background: Multi-drug resistant Gram-negative bacteria (GNB) are major contributors to the antimicrobial resistance (AMR) burden. AMR mechanisms are primarily mediated by proteoforms; therefore, proteomic analyses of GNB offers a significant advantage in understanding the mechanisms of AMR. A large portion of these mechanisms are mediated by membrane proteins; however, they are often difficult to extract due to their hydrophobic nature and complex interactions with other components of the cell membrane. To extract the greatest number of proteoforms, an efficient homogenisation protocol is required to effectively disrupt the rigid cell wall and membrane. Methods: Using Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa, we systematically compared the extraction efficiency of bead-beating with flash frozen and lyophilized cell pellets. Results: We demonstrate that lyophilization improves bead-beating extraction methods by increasing the detection of membrane proteins. We detected numerous unique membrane proteins in each bacterial isolate, including ABC transporters and proteins involved in lipopolysaccharide synthesis, when lyophilizing prior to bead-beating, compared to only flash-freezing. Conclusions: As membrane proteins play a central role in AMR mechanisms, this improvement in their isolation and identification will aid in understanding the resistance and molecular mechanisms associated with multi-drug resistant GNB. Full article

(This article belongs to the Section Proteomics Technology and Methodology Development)

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8 pages, 520 KB

Open AccessTechnical Note

An Ultrafast GPU-Enabled MGVB

by Metodi V. Metodiev

Proteomes 2026, 14(3), 34; https://doi.org/10.3390/proteomes14030034 - 7 Jul 2026

Viewed by 421

Abstract

Background: cMGVB is a graphical processing unit (GPU)-enabled implementation of the computational proteomics data analysis toolset MGVB. MGVB was released in 2025 as a Linux program designed to run on multi-node servers. It utilizes a novel algorithm for finding combinations of post-translational [...] Read more.

Background: cMGVB is a graphical processing unit (GPU)-enabled implementation of the computational proteomics data analysis toolset MGVB. MGVB was released in 2025 as a Linux program designed to run on multi-node servers. It utilizes a novel algorithm for finding combinations of post-translational modification in peptide MS/MS data. The original combinatorial algorithm required a significant amount of resources to be practical. Hence, the aim of the research reported here was to port the algorithm to GPU and thus increase its speed and efficiency. Methods: To accomplish this it was recoded in CUDA C; recursive functions and data structures were re-implemented as non-recursive, and the algorithm was incorporated in a new version of MGVB, now termed cMGVB. Results: The re-implemented algorithm is much faster and, unlike the original program, can run on single CPU workstations equipped with inexpensive GPUs and still be much faster than the original algorithm running on HPC clusters. A typical focused search is completed in about a minute by cMGVB compared to 10–15 min by the original implementation. Illustrative case studies are presented and discussed in this report. Conclusions: cMGVB enables workflows that were not practical or even possible with the original MGVB. Full article

(This article belongs to the Section Proteome Bioinformatics)

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13 pages, 3418 KB

Open AccessArticle

A Dual-Background Statistical Framework for Phosphoproteomics Highlights Intrinsic, High-Confidence Phosphorylation Signature by Mitigating Orthogonal Sources of Bias

by Bin Deng

Proteomes 2026, 14(3), 33; https://doi.org/10.3390/proteomes14030033 - 7 Jul 2026

Viewed by 509

Abstract

Background: Distinguishing genuine kinase–substrate motifs from background noise is a growing challenge, as mass spectrometry (MS)-based global phosphoproteomics identifies a rapidly expanding set of phosphorylation sites. One of the major limitations is selecting an appropriate background model that systematically controls both technical and [...] Read more.

Background: Distinguishing genuine kinase–substrate motifs from background noise is a growing challenge, as mass spectrometry (MS)-based global phosphoproteomics identifies a rapidly expanding set of phosphorylation sites. One of the major limitations is selecting an appropriate background model that systematically controls both technical and biological sources of bias. Although using the entire proteome as a background in a FASTA format considers the overall amino acid composition, it is still prone to biases from protein abundance and the uneven distribution of sequence space (particularly around low-abundance proteins). By contrast, internal background methods can control experiment-specific detection biases, but they may not fully capture residue-specific compositions or general trends in phosphorylation. Methods: I develop a Dual-Background Enrichment (DBE) framework with a position-specific enrichment (PSE) strategy, which involves analyzing motif enrichment against two distinct background models: (1) A residue-heterogeneous internal background composed of phospho-motifs centered on the residue; e.g., phosphoserine (pS) motifs are tested relative to the pool of all detected phosphothreonine (pT) and phosphotyrosine (pY) motifs from the same experiment. (2) A FASTA background that includes all S, T, and Y residues in the UniProtKB proteome sequences. Results: Motifs are classified as high confidence if they meet statistical significance (q ≤ 0.05, fold enrichment > 1.5) against both background models. Conclusion: By applying the DBE strategy to a large-scale phosphoproteomics dataset, we distinguish motifs driven by amino acid composition (enriched in FASTA background only) from those reflecting kinase substrate specificity (enriched in both backgrounds). This dual-reference approach reduces false positives arising from sequence composition bias and enriches high-confidence candidate kinase recognition motifs. Full article

(This article belongs to the Section Proteome Bioinformatics)

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Graphical abstract

24 pages, 1650 KB

Open AccessReview

A One Health Framework for Proteomics Across the Tree of Life to Advance Food Security, Animal Health, and Ecosystem Resilience

by Tarun Mishra, Ritudhwaj Tiwari, Tuyelee Das and Maneesh Lingwan

Proteomes 2026, 14(3), 32; https://doi.org/10.3390/proteomes14030032 - 24 Jun 2026

Viewed by 1760

Abstract

As global ecosystems and food systems face unprecedented anthropogenic and climatic challenges, there is a demand for an integrated understanding of biological systems. Proteomics has emerged as a definitive approach offering a direct view of the molecular phenotype, yet it is traditionally separated [...] Read more.

As global ecosystems and food systems face unprecedented anthropogenic and climatic challenges, there is a demand for an integrated understanding of biological systems. Proteomics has emerged as a definitive approach offering a direct view of the molecular phenotype, yet it is traditionally separated into plant and animal disciplines. With recent advances in mass spectrometry (MS) and bioinformatics tools, this prospective review proposes that combining a One Health proteomics approach with deep-learning data analysis can revolutionize global food security, animal productivity, and ecosystem health by uncovering proteoform signatures that drive resilience across life. The potential of a unified One Health proteomic framework, highlighting major developments, including 4D proteomics, Data-Independent Acquisition (DIA), and single-cell resolution, and emphasizes their capacity to resolve the complex proteoform landscape across kingdoms. Review emphasizes the applications of proteogenomics as a cross-disciplinary tool to improve genome annotations, explain evolutionary differences, discover biomarkers in animals and resolve complex signaling networks in plants under stress. Nevertheless, contemporary proteogenomics methods still show limitations in their ability to comprehensively resolve proteoforms due to the fact that the use of peptide-based approaches makes it difficult to fully appreciate the post-translational modifications specific to each protein isoform. We show that One Health proteomics will provide a transformative roadmap for deciphering the functional proteoform signatures that underpin resilience across the tree of life. Full article

(This article belongs to the Special Issue Plant Genomics and Proteomics)

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Proteomes, Volume 14, Issue 3 (September 2026) – 18 articles

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