BACKGROUND: It is well established that the cancerous transformation of cells is accompanied by profound alterations in glycosylation. In this study, we demonstrate the diagnostic potential of N-glycan profiling in tissue specimens from patients, primarily representing the two major types of lung cancer: non-small cell and small cell lung cancer. METHODS: Lung tissues and biopsies obtained from surgery and bronchoscopy underwent sample processing and enzymatic digestion. After labeling, glycans were analyzed employing matrix-assisted laser desorption/ionization mass spectrometry. Statistical analysis was conducted using methods following principles of compositional data analysis. RESULTS: Comparison of glycan profiles demonstrated an increase in paucimannose and high mannose glycans in most tumor specimens, including those with inflammation and histological negative for malignancy. Cancerous tissues exhibited more profound changes in glycosylation. Despite the high heterogeneity in profiles, two main groups of not detected glycans in peritumoral tissues, considered as controls, were observed to correlate with cancer progression in patients. One with complex polylactosamine multifucosylated glycans frequently harboring terminal N-acetyl-glucosamine residues. These glycans were present in most tumors, with their numbers and intensities increasing as cancer progressed. In contrast, the second group exhibited polylactosamine glycans sporadically. Instead, the biopsies of several patients with rapid progression displayed a significant presence in a set of tri- and tetra-antennary core fucosylated glycans having mostly unoccupied N-acetyl-glucosamine residues unless carrying additional fucose unit(s). CONCLUSIONS: The results imply distinct glycosylation patterns even in patients with the same histological type of lung cancer, supporting trends toward personalized diagnosis and more tailored therapies. Currently, tissue biopsies remain the gold standard for diagnosing premalignant and malignant lesions in the lung. Expanded knowledge on glycosylation in these lesions could contribute to improved diagnostic accuracy and better monitoring of malignant disease progression in clinical practice.
- Klíčová slova
- N-glycans, Branched glycans, Fucosylation, Glycosylation, Lung cancer, MALDI-MS,
- MeSH
- glykosylace MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádory plic * diagnóza metabolismus patologie MeSH
- nemalobuněčný karcinom plic diagnóza metabolismus patologie MeSH
- plíce * patologie metabolismus MeSH
- polysacharidy * metabolismus chemie MeSH
- prognóza MeSH
- prospektivní studie MeSH
- senioři MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- polysacharidy * MeSH
High-risk human papillomaviruses (HPVs) cause various cancers. While type-specific prophylactic vaccines are available, additional anti-viral strategies are highly desirable. Initial HPV cell entry involves receptor-switching induced by structural capsid modifications. These modifications are initiated by interactions with cellular heparan sulphates (HS), however, their molecular nature and functional consequences remain elusive. Combining virological assays with hydrogen/deuterium exchange mass spectrometry, and atomic force microscopy, we investigate the effect of capsid-HS binding and structural activation. We show how HS-induced structural activation requires a minimal HS-chain length and simultaneous engagement of several binding sites by a single HS molecule. This engagement introduces a pincer-like force that stabilizes the capsid in a conformation with extended capsomer linkers. It results in capsid enlargement and softening, thereby likely facilitating L1 proteolytic cleavage and subsequent L2-externalization, as needed for cell entry. Our data supports the further devising of prophylactic strategies against HPV infections.
- MeSH
- heparitinsulfát * metabolismus chemie MeSH
- infekce papilomavirem virologie MeSH
- internalizace viru * MeSH
- kapsida * metabolismus chemie MeSH
- lidé MeSH
- lidské papilomaviry MeSH
- lidský papilomavirus 16 metabolismus fyziologie MeSH
- mikroskopie atomárních sil * MeSH
- Papillomaviridae fyziologie MeSH
- polysacharidy metabolismus chemie MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- virové plášťové proteiny * metabolismus chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- heparitinsulfát * MeSH
- polysacharidy MeSH
- virové plášťové proteiny * MeSH
Dynamic changes in protein glycosylation impact human health and disease progression. However, current resources that capture disease and phenotype information focus primarily on the macromolecules within the central dogma of molecular biology (DNA, RNA, proteins). To gain a better understanding of organisms, there is a need to capture the functional impact of glycans and glycosylation on biological processes. A workshop titled "Functional impact of glycans and their curation" was held in conjunction with the 16th Annual International Biocuration Conference to discuss ongoing worldwide activities related to glycan function curation. This workshop brought together subject matter experts, tool developers, and biocurators from over 20 projects and bioinformatics resources. Participants discussed four key topics for each of their resources: (i) how they curate glycan function-related data from publications and other sources, (ii) what type of data they would like to acquire, (iii) what data they currently have, and (iv) what standards they use. Their answers contributed input that provided a comprehensive overview of state-of-the-art glycan function curation and annotations. This report summarizes the outcome of discussions, including potential solutions and areas where curators, data wranglers, and text mining experts can collaborate to address current gaps in glycan and glycosylation annotations, leveraging each other's work to improve their respective resources and encourage impactful data sharing among resources. Database URL: https://wiki.glygen.org/Glycan_Function_Workshop_2023.
- MeSH
- biokurátorství MeSH
- datové kurátorství * metody MeSH
- glykosylace MeSH
- lidé MeSH
- polysacharidy * metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- kongresy MeSH
- Research Support, N.I.H., Extramural MeSH
- Geografické názvy
- Itálie MeSH
- Názvy látek
- polysacharidy * MeSH
Mammalian spermatozoa have a surface covered with glycocalyx, consisting of heterogeneous glycoproteins and glycolipids. This complexity arises from diverse monosaccharides, distinct linkages, various isomeric glycans, branching levels, and saccharide sequences. The glycocalyx is synthesized by spermatozoa developing in the testis, and its subsequent alterations during their transit through the epididymis are a critical process for the sperm acquisition of fertilizing ability. In this study, we performed detailed analysis of the glycocalyx on the sperm surface of bull spermatozoa in relation to individual parts of the epididymis using a wide range (24) of lectins with specific carbohydrate binding preferences. Fluorescence analysis of intact sperm isolated from the bull epididymides was complemented by Western blot detection of protein extracts from the sperm plasma membrane fractions. Our experimental results revealed predominant sequential modification of bull sperm glycans with N-acetyllactosamine (LacNAc), followed by subsequent sialylation and fucosylation in a highly specific manner. Additionally, variations in the lectin detection on the sperm surface may indicate the acquisition or release of glycans or glycoproteins. Our study is the first to provide a complex analysis of the bull sperm glycocalyx modification during epididymal maturation.
- Klíčová slova
- cattle, epididymis, glycan, lectin, plasma membrane, sperm surface, spermatozoa,
- MeSH
- epididymis * metabolismus cytologie MeSH
- glykokalyx * metabolismus MeSH
- glykoproteiny metabolismus MeSH
- lektiny * metabolismus MeSH
- polysacharidy metabolismus MeSH
- skot MeSH
- spermie * metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- glykoproteiny MeSH
- lektiny * MeSH
- polysacharidy MeSH
Prostate cancer (PCa) is the second most common cancer. In this paper, the isolation and properties of exosomes as potential novel liquid biopsy markers for early PCa liquid biopsy diagnosis are investigated using two prostate human cell lines, i.e., benign (control) cell line RWPE1 and carcinoma cell line 22Rv1. Exosomes produced by both cell lines are characterised by various methods including nanoparticle-tracking analysis, dynamic light scattering, scanning electron microscopy and atomic force microscopy. In addition, surface plasmon resonance (SPR) is used to study three different receptors on the exosomal surface (CD63, CD81 and prostate-specific membrane antigen-PMSA), implementing monoclonal antibodies and identifying the type of glycans present on the surface of exosomes using lectins (glycan-recognising proteins). Electrochemical analysis is used to understand the interfacial properties of exosomes. The results indicate that cancerous exosomes are smaller, are produced at higher concentrations, and exhibit more nega tive zeta potential than the control exosomes. The SPR experiments confirm that negatively charged α-2,3- and α-2,6-sialic acid-containing glycans are found in greater abundance on carcinoma exosomes, whereas bisecting and branched glycans are more abundant in the control exosomes. The SPR results also show that a sandwich antibody/exosomes/lectins configuration could be constructed for effective glycoprofiling of exosomes as a novel liquid biopsy marker.
- Klíčová slova
- exosomes, microscopy techniques, nanoparticle tracking analysis, prostate cancer, self-assembled monolayer, surface plasmon resonance,
- MeSH
- exozómy * chemie MeSH
- karcinom * metabolismus patologie MeSH
- lektiny analýza metabolismus MeSH
- lidé MeSH
- polysacharidy analýza metabolismus MeSH
- tekutá biopsie MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- lektiny MeSH
- polysacharidy MeSH
Breast cancer is a highly heterogeneous disease. Its intrinsic subtype classification for diagnosis and choice of therapy traditionally relies on the presence of characteristic receptors. Unfortunately, this classification is often not sufficient for precise prediction of disease prognosis and treatment efficacy. The N-glycan profiles of 145 tumors and 10 healthy breast tissues were determined using Matrix-Assisted Laser Desorption-Ionization Time-of-Flight Mass Spectrometry. The tumor samples were classified into Mucinous, Lobular, No-Special-Type, Human Epidermal Growth Factor 2 + , and Triple-Negative Breast Cancer subtypes. Statistical analysis was conducted using the reproducibility-optimized test statistic software package in R, and the Wilcoxon rank sum test with continuity correction. In total, 92 N-glycans were detected and quantified, with 59 consistently observed in over half of the samples. Significant variations in N-glycan signals were found among subtypes. Mucinous tumor samples exhibited the most distinct changes, with 28 significantly altered N-glycan signals. Increased levels of tri- and tetra-antennary N-glycans were notably present in this subtype. Triple-Negative Breast Cancer showed more N-glycans with additional mannose units, a factor associated with cancer progression. Individual N-glycans differentiated Human Epidermal Growth Factor 2 + , No-Special-Type, and Lobular cancers, whereas lower fucosylation and branching levels were found in N-glycans significantly increased in Luminal subtypes (Lobular and No-Special-Type tumors). Clinically normal breast tissues featured a higher abundance of signals corresponding to N-glycans with bisecting moiety. This research confirms that histologically distinct breast cancer subtypes have a quantitatively unique set of N-glycans linked to clinical parameters like tumor size, proliferative rate, lymphovascular invasion, and metastases to lymph nodes. The presented results provide novel information that N-glycan profiling could accurately classify human breast cancer samples, offer stratification of patients, and ongoing disease monitoring.
- MeSH
- epidermální růstové faktory MeSH
- lidé MeSH
- polysacharidy metabolismus MeSH
- prognóza MeSH
- reprodukovatelnost výsledků MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- triple-negativní karcinom prsu * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- epidermální růstové faktory MeSH
- polysacharidy MeSH
Stem bending in trees induces flexure wood but its properties and development are poorly understood. Here, we investigated the effects of low-intensity multidirectional stem flexing on growth and wood properties of hybrid aspen, and on its transcriptomic and hormonal responses. Glasshouse-grown trees were either kept stationary or subjected to several daily shakes for 5 wk, after which the transcriptomes and hormones were analyzed in the cambial region and developing wood tissues, and the wood properties were analyzed by physical, chemical and microscopy techniques. Shaking increased primary and secondary growth and altered wood differentiation by stimulating gelatinous-fiber formation, reducing secondary wall thickness, changing matrix polysaccharides and increasing cellulose, G- and H-lignin contents, cell wall porosity and saccharification yields. Wood-forming tissues exhibited elevated jasmonate, polyamine, ethylene and brassinosteroids and reduced abscisic acid and gibberellin signaling. Transcriptional responses resembled those during tension wood formation but not opposite wood formation and revealed several thigmomorphogenesis-related genes as well as novel gene networks including FLA and XTH genes encoding plasma membrane-bound proteins. Low-intensity stem flexing stimulates growth and induces wood having improved biorefinery properties through molecular and hormonal pathways similar to thigmomorphogenesis in herbaceous plants and largely overlapping with the tension wood program of hardwoods.
- Klíčová slova
- Populus tremula × tremuloides, flexure wood, jasmonic acid signaling, mechanostimulation, polyamines, saccharification, thigmomorphogenesis, wood development,
- MeSH
- buněčná stěna metabolismus MeSH
- celulosa metabolismus MeSH
- dřevo * MeSH
- polyaminy analýza metabolismus farmakologie MeSH
- polysacharidy metabolismus MeSH
- Populus * genetika MeSH
- regulace genové exprese u rostlin MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- celulosa MeSH
- jasmonic acid MeSH Prohlížeč
- polyaminy MeSH
- polysacharidy MeSH
Protein glycosylation is one of the most common PTMs and many cell surface receptors, extracellular proteins, and biopharmaceuticals are glycosylated. However, HDX-MS analysis of such important glycoproteins has so far been limited by difficulties in determining the HDX of the protein segments that contain glycans. We have developed a column containing immobilized PNGase Rc (from Rudaea cellulosilytica) that can readily be implemented into a conventional HDX-MS setup to allow improved analysis of glycoproteins. We show that HDX-MS with the PNGase Rc column enables efficient online removal of N-linked glycans and the determination of the HDX of glycosylated regions in several complex glycoproteins. Additionally, we use the PNGase Rc column to perform a comprehensive HDX-MS mapping of the binding epitope of a mAb to c-Met, a complex glycoprotein drug target. Importantly, the column retains high activity in the presence of common quench-buffer additives like TCEP and urea and performed consistent across 114 days of extensive use. Overall, our work shows that HDX-MS with the integrated PNGase Rc column can enable fast and efficient online deglycosylation at harsh quench conditions to provide comprehensive analysis of complex glycoproteins.
- MeSH
- glykopeptidasa MeSH
- glykoproteiny * analýza MeSH
- glykosylace MeSH
- polysacharidy * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- glykopeptidasa MeSH
- glykoproteiny * MeSH
- polysacharidy * MeSH
Galectins are proteins of the family of human lectins. By binding terminal galactose units of cell surface glycans, they moderate biological and pathological processes such as cell signaling, cell adhesion, apoptosis, fibrosis, carcinogenesis, and metabolic disorders. The binding of monovalent glycans to galectins is usually relatively weak. Therefore, the presentation of carbohydrate ligands on multivalent scaffolds can efficiently increase and/or discriminate the affinity of the glycoconjugate to different galectins. A library of glycoclusters and glycodendrimers with various structural presentations of the common functionalized N-acetyllactosamine ligand was prepared to evaluate how the mode of presentation affects the affinity and selectivity to the two most abundant galectins, galectin-1 (Gal-1) and galectin-3 (Gal-3). In addition, the effect of a one- to two-unit carbohydrate spacer on the affinity of the glycoconjugates was determined. A new design of the biolayer interferometry (BLI) method with specific AVI-tagged constructs was used to determine the affinity to galectins, and compared with the gold-standard method of isothermal titration calorimetry (ITC). This study reveals new routes to low nanomolar glycoconjugate inhibitors of galectins of interest for biomedical research.
- Klíčová slova
- Biolayer interferometry, Carbohydrate, Click chemistry, Galectin, Glycoconjugate, Multivalency, Transglycosylation,
- MeSH
- galektiny * metabolismus MeSH
- glykokonjugáty * farmakologie chemie MeSH
- lidé MeSH
- ligandy MeSH
- polysacharidy metabolismus MeSH
- sacharidy chemie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- galektiny * MeSH
- glykokonjugáty * MeSH
- ligandy MeSH
- polysacharidy MeSH
- sacharidy MeSH
Polyglutamic acid (PGA), a protein in the mucilage of PGA-producing Bacillus spp., has expected applications in medical and biotechnological industries. Although the Bacillaceae family contains over 100 genera, research on bacterial PGA has exclusively focused on the genus Bacillus, especially B. subtilis var. natto and B. licheniformis. In the present study, indigenous Bacillaceae family strains were isolated from withered leaves and soil samples and screened for PGA production. As a result of the screening, the strain 8h was found to produce a mucilage possessing greater viscosity than PGA of B. subtilis var. natto (natto PGA). Biochemical analyses revealed that the 8h mucilage contains 63% protein and 37% polysaccharide, while mucilage of B. subtilis var. natto is composed of 61% protein and 39% polysaccharide. The most plentiful amino acid in 8h mucilage protein was glutamate (43%, mol/mol), which is similar to that of natto PGA, suggesting that it possesses characteristics of PGA. Although natto mucilage contains fructan, glucan was found as the polysaccharide of 8h mucilage. While phylogenetic studies indicated that the strain 8h belongs to Peribacillus simplex, the yield of the viscous mucilage by strain 8h was significantly higher than P. simplex type strain, suggesting that 8h is a mucilage-overproducing strain of P. simplex. Interestingly, 8h mucilage protein was found to contain more hydrophobic amino acid residues than natto PGA, suggesting that its amphiphilicity is suitable as a drug carrier and adjuvant. The present study is the first report of viscous mucilage and PGA-like protein produced by the genus Peribacillus.
- Klíčová slova
- Bacillaceae, Mucilage protein, Peribacillus simplex, Polyglutamic acid, Soil bacteria,
- MeSH
- Bacillus subtilis metabolismus MeSH
- Bacillus * metabolismus MeSH
- fylogeneze MeSH
- kyselina polyglutamová * analýza metabolismus MeSH
- polysacharidy metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- kyselina polyglutamová * MeSH
- polysacharidy MeSH
