Showing papers on "Glycome published in 2014"
TL;DR: This work has shown that glycan modifications of intracellular and extracellular proteins have critical functions in almost all biological pathways and is providing insights into how glycans function in recognition and signaling within an organism and with microbes and pathogens.
316 citations
TL;DR: This study has revealed very extensive and complex changes in IgG glycosylation with age, and the combined index composed of only three glycans explained up to 58% of variance in age, considerably more than other biomarkers of age like telomere lengths.
Abstract: Aging is a complex process of accumulation of molecular, cellular, and organ damage, leading to loss of function and increased vulnerability to disease and finally to death (1). It is well known that lifestyle choices such as smoking and physical activity can hasten or delay the aging process (2). Such observations have led to the search for molecular markers of age that can be used to predict, monitor, and provide insight into age-associated physiological decline and disease.
Protein structure is defined by the sequence of nucleotides in the corresponding genes, thus the polypeptide sequence of a protein cannot change with age. However, an important structural and functional element of the majority of proteins are the glycans that participate in virtually all physiological processes (3). Glycans are product of a complex pathway that involves hundreds of different proteins and are encoded in a complex dynamic network of hundreds of genes (4). Epigenetic regulation of gene expression is expected to affect protein glycosylation and several publications recently reported this effect (5–8). Changes in glycosylation with age have been shown over 20 years ago (9) and have also replicated in recent large population studies (10–13).
Immunoglobulin G (IgG) is an excellent model glycoprotein because its glycosylation has been well defined (Figure 1), and many important functional effects of alternative IgG glycosylation have been described (14). For example, glycosylation acts as a switch between pro- and anti-inflammatory IgG functionality. Most of the IgG molecules are not sialylated and are proinflammatory. Terminal α2,6-sialylation of IgG glycans decreases the ability of IgG to bind to activating FcγRs and promotes recognition by DC-SIGN, which increases expression of inhibitory FcγRIIB and is anti-inflammatory (15). Another fascinating example is the role of core fucose in the modulation of antibody-dependent cellular cytotoxicity: IgG-containing glycans that lack core fucose have 100-fold increased affinity for FcγRIIIA and are therefore much more efficient in activating antibody-dependent cellular cytotoxicity than fucosylated glycoforms of the same molecule (16). On average, 95% of the IgG population is core fucosylated (12); thus, most of the immunoglobulins have a “safety switch,” which prevents them from activating antibody-dependent cellular cytotoxicity. Malfunction of this system appears to be associated with autoimmune diseases as indicated by both pleiotropic effects of genes that associate with IgG glycosylation on different inflammatory and autoimmune diseases, and the observed alterations in IgG glycosylation in systemic lupus erythematous (17) and many inflammatory diseases (18).
Figure 1.
UPLC analysis of immunoglobulin G (IgG) glycosylation. Each IgG contains one conserved N-glycosylation site on Asn197 of its heavy chain. Different glycans can be attached to this site and the process seems to be highly regulated. UPLC analysis can reveal ...
Interindividual variability of IgG glycosylation in a population is large (12) and it appears to be affected by both variation in DNA sequence (19) and environmental factors (11). Most of the studies that investigated glycosylation changes with age were either of limited size or were performed on the total plasma glycome; thus, in addition to changes in glycosylation, the observed differences reflected changes in the concentration of individual plasma proteins. In this study, we focused on glycosylation of IgG and analyzed more than 5,000 individuals from four different European populations to provide definitive data about changes in IgG glycosylation through the lifetime.
260 citations
TL;DR: To investigate the potential role of HMGs as decoy receptors for rotavirus (RV), a leading cause of severe gastroenteritis in children, and indicate that sialic acid is not required for binding of glycans to individual VP8* domains, the functional glycome of human milk was explored using shotgun glycomics.
107 citations
TL;DR: Using this framework, 103 highly confident N-linked glycopeptides from 53 sites across 33 glycoproteins were identified in complex human serum proteome samples using conventional proteomic platforms with standard depletion of the 7-most abundant proteins, indicating that the method is ready to be used for characterizing site-specific protein glycosylation in complex samples.
Abstract: Glycosylation is an important protein modification that involves enzymatic attachment of sugars to amino acid residues. Understanding the structure of these sugars and the effects of glycosylation are vital for developing indicators of disease development and progression. Although computational methods based on mass spectrometric data have proven to be effective in monitoring changes in the glycome, developing such methods for the glycoproteome are challenging, largely due to the inherent complexity in simultaneously studying glycan structures with their corresponding glycosylation sites. This paper introduces a computational framework for identifying intact N-linked glycopeptides, i.e. glycopeptides with N-linked glycans attached to their glycosylation sites, in complex proteome samples. Scoring algorithms are presented for tandem mass spectra of glycopeptides resulting from collision-induced dissociation (CID), higher-energy C-trap dissociation (HCD), and electron transfer dissociation (ETD) fragmentati...
89 citations
TL;DR: The sensitivity of the analysis system and the provision for generating 'designer' microarrays from glycomes to identify novel ligands of biological relevance are highlighted in this review.
75 citations
TL;DR: In this paper, a unique system-based approach was proposed to identify connections between microRNA and the glycome, revealing critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and glycome.
Abstract: Cell surface glycans form a critical interface with the biological milieu, informing diverse processes from the inflammatory cascade to cellular migration. Assembly of discrete carbohydrate structures requires the coordinated activity of a repertoire of proteins, including glycosyltransferases and glycosidases. Little is known about the regulatory networks controlling this complex biosynthetic process. Recent work points to a role for microRNA (miRNA) in the regulation of specific glycan biosynthetic enzymes. Herein we take a unique systems-based approach to identify connections between miRNA and the glycome. By using our glycomic analysis platform, lectin microarrays, we identify glycosylation signatures in the NCI-60 cell panel that point to the glycome as a direct output of genomic information flow. Integrating our glycomic dataset with miRNA data, we map miRNA regulators onto genes in glycan biosynthetic pathways (glycogenes) that generate the observed glycan structures. We validate three of these predicted miRNA/glycogene regulatory networks: high mannose, fucose, and terminal β-GalNAc, identifying miRNA regulation that would not have been observed by traditional bioinformatic methods. Overall, our work reveals critical nodes in the global glycosylation network accessible to miRNA regulation, providing a bridge between miRNA-mediated control of cell phenotype and the glycome.
68 citations
TL;DR: Compared with the labeling methods using free azidosugars, this method offers improved labeling efficiency and high specificity and should facilitate the elucidation of the functional role of glycans in cancer biology.
Abstract: Although it has been well known that dynamic changes in glycosylation are associated with tumor progression, it remains challenging to selectively visualize the cancer glycome in vivo Herein, a strategy for the targeted imaging of tumor-associated glycans by using ligand-targeted liposomes encapsulating azidosugars is described The intravenously injected liposomal nanoparticles selectively bound to the cancer-cell-specific receptors and installed azides into the melanoma glycans in a xenograft mouse model in a tissue-specific manner Subsequently, a copper-free click reaction was performed in vivo to chemoselectively conjugate the azides with a near-infrared fluorescent dye The glycosylation dynamics during tumor growth were monitored by in vivo fluorescence imaging Furthermore, the newly synthesized sialylated glycoproteins were enriched during tumor growth and identified by glycoproteomics Compared with the labeling methods using free azidosugars, this method offers improved labeling efficiency and high specificity and should facilitate the elucidation of the functional role of glycans in cancer biology
63 citations
TL;DR: Epigenetic recording of acquired characteristics and their transgenerational inheritance could be important mechanisms used by higher organisms to compete or collaborate with microorganisms.
62 citations
TL;DR: A chemical approach for analyzing the dynamic cardiac glycome by metabolically labeling the cardiac glycans with azidosugars in living rats, which revealed an increase of sialic acid biosynthesis upon the induction of cardiac hypertrophy.
Abstract: In the heart, glycosylation is involved in a variety of physiological and pathological processes. Cardiac glycosylation is dynamically regulated, which remains challenging to monitor in vivo. Here we describe a chemical approach for analyzing the dynamic cardiac glycome by metabolically labeling the cardiac glycans with azidosugars in living rats. The azides, serving as a chemical reporter, are chemoselectively conjugated with fluorophores using copper-free click chemistry for glycan imaging; derivatizing azides with affinity tags allows enrichment and proteomic identification of glycosylated cardiac proteins. We demonstrated this methodology by visualization of the cardiac sialylated glycans in intact hearts and identification of more than 200 cardiac proteins modified with sialic acids. We further applied this methodology to investigate the sialylation in hypertrophic hearts. The imaging results revealed an increase of sialic acid biosynthesis upon the induction of cardiac hypertrophy. Quantitative prot...
62 citations
TL;DR: This review focuses on a higher level of glycan organization, the formation of clustered saccharide patches (CSPs), which can constitute unique ligands for highly specific interactions, and presents a wealth of evidence for CSPs-mediated interactions.
Abstract: All cells in nature are covered with a dense and complex array of glycan chains. Specific recognition and binding of glycans is a critical aspect of cellular interactions, both within and between species. Glycan–protein interactions tend to be of low affinity but high specificity, typically utilizing multivalency to generate the affinity required for biologically relevant binding. This review focuses on a higher level of glycan organization, the formation of clustered saccharide patches (CSPs), which can constitute unique ligands for highly specific interactions. Due to technical challenges, this aspect of glycan recognition remains poorly understood. We present a wealth of evidence for CSPs-mediated interactions, and discuss recent advances in experimental tools that are beginning to provide new insights into the composition and organization of CSPs. The examples presented here are likely the tip of the iceberg, and much further work is needed to elucidate fully this higher level of glycan organization.
59 citations
TL;DR: The results reveal F77 antigen to be expressed on blood group H on a 6-linked branch of a poly-N-acetyllactosamine backbone and show that mAb F77 can also bind to blood group A and B analogs but with lower intensities.
TL;DR: Human cells have developed mechanisms to simultaneously and reproducibly generate subcellular-specific N-glycosylation using a shared biosynthetic machinery.
Abstract: Glycoproteins perform extra- and intracellular functions in innate and adaptive immunity by lectin-based interactions to exposed glyco-determinants. Herein, we document and mechanistically explain the formation of subcellular-specific N-glycosylation determinants on glycoproteins trafficking through the shared biosynthetic machinery of human cells. LC-MS/MS-based quantitative glycomics showed that the secreted glycoproteins of eight human breast epithelial cells displaying diverse geno- and phenotypes consistently displayed more processed, primarily complex type, N-glycans than the high mannose-rich microsomal glycoproteins. Detailed subcellular glycome profiling of proteins derived from three breast cell lines (MCF7/MDA468/MCF10A) demonstrated that secreted glycoproteins displayed significantly more α-sialylation and α1,6-fucosylation, but less α-mannosylation, than both the intermediately glycan-processed cell surface glycoproteomes and the under-processed microsomal glycoproteomes. Subcellular proteomics and gene ontology revealed substantial presence of endoplasmic reticulum resident glycoproteins in the microsomes and confirmed significant enrichment of secreted and cell surface glycoproteins in the respective subcellular fractions. The solvent accessibility of the glycosylation sites on maturely folded proteins of the 100 most abundant putative N-glycoproteins observed uniquely in the three subcellular glycoproteomes correlated with the glycan type processing thereby mechanistically explaining the formation of subcellular-specific N-glycosylation. In conclusion, human cells have developed mechanisms to simultaneously and reproducibly generate subcellular-specific N-glycosylation using a shared biosynthetic machinery. This aspect of protein-specific glycosylation is important for structural and functional glycobiology and discussed here in the context of the spatio-temporal interaction of glyco-determinants with lectins central to infection and immunity.
TL;DR: Altered glycosylation profiles both for total plasma glycoproteins and on individual proteins during pregnancy are shown, which may contribute to immunosuppression and have other biological functions.
Abstract: During pregnancy, the mother faces a major immunological challenge. Most of the major plasma proteins have important immunological functions, and altered levels of these major proteins have been reported during pregnancy, potentially providing immunosuppression. A large number of the high abundance plasma proteins are post-translationally modified by N-glycans, and while it is now understood that these glycans may also affect the immunological functions, their pattern has not been studied in relation to pregnancy. Here, the N-glycosylation profile of 32 pregnant women was determined over the course of their pregnancy using a multiplexed CGE-LIF method. Moreover, for 6 women, the glycosylation profiles of the proteins IgG, IgA, and alpha1-antitrypsin were monitored. For total plasma, 16 glycan signals showed differential expression during pregnancy. In general the levels of largely sialylated bi-, tri-, and tetra-antennary glycans were increased during pregnancy, while biantennary glycans with no more than one sialic acid were decreased. Similarly altered glycosylation profiles were observed for the individual proteins IgG, with a decrease of digalactosylated biantennary glycans after delivery, and alpha1-antitrypsin, on which increased levels of triantennary glycans were observed during pregnancy. Overall, these results show altered glycosylation profiles both for total plasma glycoproteins and on individual proteins during pregnancy, which may contribute to immunosuppression and have other biological functions.
TL;DR: This work has developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells and directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N- glycans.
Abstract: Cell surfaces are covered with a dense carbohydrate layer referred to as the glycocalyx. Because different cell types express different glycan signatures, it is of paramount importance to have robust methods to analyze the glycome of living cells. To achieve this, a common procedure involves cell lysis and extraction of membrane (glyco)proteins and yields a major proportion of high-mannose N-glycans that most likely stem from intracellular proteins derived from the ER. Using HEK 293 cells as a model system, we developed a reproducible, sensitive, and fast method to profile surface N-glycosylation from living cells. We directly released glycopeptides from cell surfaces through tryptic digestion of freshly harvested and vital cells, thereby improving the detection and quantification of complex-type N-glycans by increasing their relative amount from 14 to 85%. It was also possible to detect 25 additional structures in HEK 293, 48 in AGE1.HN, 42 in CHO-K1, and 51 in Hep G2 cells. The additional signals provided deeper insight into cell-type-specific N-glycan features such as antennarity, fucosylation, and sialylation. Thus, this protocol, which can potentially be applied to any cells, will be useful in the fields of glycobiotechnology and biomarker discovery.
TL;DR: Using carbon-LC-ESI-CID-MS/MS of protein released native N-glycans, Wang et al. as mentioned in this paper accurately profiled the secretome Nglycosylation of six human epithelial breast cells.
Abstract: The secreted cellular sub-proteome (secretome) is a rich source of biologically active glycoproteins. N-Glycan profiling of secretomes of cultured cancer cells provides an opportunity to investigate the link between protein N-glycosylation and tumorigenesis. Utilizing carbon-LC–ESI-CID-MS/MS of protein released native N-glycans, we accurately profiled the secretome N-glycosylation of six human epithelial breast cells including normal mammary epithelial cells (HMEC) and breast cancer cells belonging to luminal A subtype (MCF7), HER2-overexpressing subtype (SKBR3), and basal B subtype (MDA-MB157, MDA-MB231, HS578T). On the basis of intact molecular mass, LC retention time, and MS/MS fragmentation, a total of 74 N-glycans were confidently identified and quantified. The secretomes comprised significant levels of highly sialylated and fucosylated complex type N-glycans, which were elevated in all cancer cells relative to HMEC (57.7–87.2% vs 24.9%, p < 0.0001 and 57.1–78.0% vs 38.4%, p < 0.0001–0.001, respectiv...
TL;DR: Characterization of the total N-linked glycome of normal, IPMN, and adenocarcinoma ductal fluid clustered samples into three discrete groups based on the prevalence of six dominant glycans and indicated that glycosylation diversity across human subjects can be reduced to simpler clusters of individuals whose N- linked glycans share structural features.
Abstract: Sensitive and specific biomarkers for pancreatic cancer are currently unavailable. The high mortality associated with adenocarcinoma of the pancreatic epithelium justifies the broadest possible search for new biomarkers that can facilitate early detection or monitor treatment efficacy. Protein glycosylation is altered in many cancers, leading many to propose that glycoproteomic changes may provide suitable biomarkers. In order to assess this possibility for pancreatic cancer, we have performed an in-depth LC-MS/MS analysis of the proteome and MS(n)-based characterization of the N-linked glycome of a small set of pancreatic ductal fluid obtained from normal, pancreatitis, intraductal papillary mucinous neoplasm (IPMN), and pancreatic adenocarcinoma patients. Our results identify a set of seven proteins that were consistently increased in cancer ductal fluid compared to normal (AMYP, PRSS1, GP2-1, CCDC132, REG1A, REG1B, and REG3A) and one protein that was consistently decreased (LIPR2). These proteins are all directly or indirectly associated with the secretory pathway in normal pancreatic cells. Validation of these changes in abundance by Western blotting revealed increased REG protein glycoform diversity in cancer. Characterization of the total N-linked glycome of normal, IPMN, and adenocarcinoma ductal fluid clustered samples into three discrete groups based on the prevalence of six dominant glycans. Within each group, the profiles of less prevalent glycans were able to distinguish normal from cancer on this small set of samples. Our results emphasize that individual variation in protein glycosylation must be considered when assessing the value of a glycoproteomic marker, but also indicate that glycosylation diversity across human subjects can be reduced to simpler clusters of individuals whose N-linked glycans share structural features.
Journal Article•
TL;DR: Complementing proteome and lipid profiling, N-glycome mapping yields important pieces of structural information to help understand the biomolecular deregulation in breast cancer development and progression, knowledge that may facilitate the discovery of candidate cancer markers and potential drug targets.
TL;DR: The new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry and allows the synthesis of biochemical reaction networks using mass spectrometry (MS) data.
Abstract: Glycosylation is among the most common and complex post-translational modifications identified to date. It proceeds through the catalytic action of multiple enzyme families that include the glycosyltransferases that add monosaccharides to growing glycans, and glycosidases which remove sugar residues to trim glycans. The expression level and specificity of these enzymes, in part, regulate the glycan distribution or glycome of specific cell/tissue systems. Currently, there is no systematic method to describe the enzymes and cellular reaction networks that catalyze glycosylation. To address this limitation, we present a streamlined machine-readable definition for the glycosylating enzymes and additional methodologies to construct and analyze glycosylation reaction networks. In this computational framework, the enzyme class is systematically designed to store detailed specificity data such as enzymatic functional group, linkage and substrate specificity. The new classes and their associated functions enable both single-reaction inference and automated full network reconstruction, when given a list of reactants and/or products along with the enzymes present in the system. In addition, graph theory is used to support functions that map the connectivity between two or more species in a network, and that generate subset models to identify rate-limiting steps regulating glycan biosynthesis. Finally, this framework allows the synthesis of biochemical reaction networks using mass spectrometry (MS) data. The features described above are illustrated using three case studies that examine: i) O-linked glycan biosynthesis during the construction of functional selectin-ligands; ii) automated N-linked glycosylation pathway construction; and iii) the handling and analysis of glycomics based MS data. Overall, the new computational framework enables automated glycosylation network model construction and analysis by integrating knowledge of glycan structure and enzyme biochemistry. All the implemented features are provided as part of the Glycosylation Network Analysis Toolbox (GNAT), an open-source, platform-independent, MATLAB based toolbox for studies of Systems Glycobiology.
TL;DR: The current state of information in regard to the schistosome glycogenome and glycome is described and the important classes of glycans and glycogenes that may be important in their generation are highlighted.
Abstract: Schistosoma mansoni and other Schistosoma sp. are multicellular parasitic helminths (worms) that infect humans and mammals worldwide. Infection by these parasites, which results in developmental maturation and sexual differentiation over a period of five to six weeks, induces antibodies to glycan antigens expressed in surface and secreted glycoproteins and glycolipids. There is growing interest in defining these unusual parasite-synthesized glycan antigens and using them to understand immune responses, their roles in immunomodulation, and in using glycan antigens as potential vaccine targets. A key problem in this area, however, has been the lack of information about the enzymes involved in elaborating the complex repertoire of glycans represented by the schistosome glycome. Recent availability of the nuclear genome sequences for Schistosoma sp. has created the opportunity to define the glycogenome, which represents the specific genes and cognate enzymes that generate the glycome. Here we describe the current state of information in regard to the schistosome glycogenome and glycome and highlight the important classes of glycans and glycogenes that may be important in their generation.
TL;DR: This work overlay predicted miRNA regulation of glycosylation related genes (glycogenes) onto maps of the common N-linked and O-linked glycan biosynthetic pathways to identify key regulatory nodes of the glycome and provides insights into glycan regulation.
TL;DR: It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC) and it was demonstrated that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucOSylated tetraanennary N-glycans were statistically increased in EOC patients.
Abstract: Alterations in glycosylation have been observed in many human diseases and specific changes in glycosylation have been proposed as relevant diagnostic information. Capillary electrophoresis coupled with laser-induced fluorescence (CE-LIF) is a robust method to quantify desialylated N-glycans that are labeled with 8-aminopyrene-1,3,6-trisulfonic acid prior to analysis. To date, only a maximum of 12 glycan structures, the most abundant ones, have been identified by CE-LIF to characterize glycome modulations of total serum in the course of the diseases. In most forms of cancer, findings using CE-LIF were limited to the increase of triantennary structures carrying a Lewisx epitope. In this work, we identified 32 linkage and positional glycan isomers in healthy human serum using exoglycosidase digestions as well as standard glycoproteins, for which we report the assignment of novel structures. It was possible to identify and quantify 34 glycan isomers in the serum of primary epithelial ovarian cancer patients (EOC). Reduced levels of diantennary structures and of high-mannose 5 were statistically significant in the EOC samples, and also, elevated branching as well as increased antennary fucosylation were observed. For the first time, we could demonstrate that not only antennary fucosylation was of relevance in tetraantennary structures but also core-fucosylated tetraantennary N-glycans were statistically increased in EOC patients. The results of the current study provide an improved dataset to be used in glycan biomarker discovery.
TL;DR: A strong correlation between differentially expressed proteins and clinical status of the patient is observed and this striking result demonstrates the potential for significant protein glycosylation alterations in clear cell renal cell carcinoma cancer plasma.
Abstract: Clear cell renal cell carcinoma is the most prevalent of all reported kidney cancer cases, and currently there are no markers for early diagnosis. This has stimulated great research interest recently because early detection of the disease can significantly improve the low survival rate. Combining the proteome, glycoproteome, and N-glycome data from clear cell renal cell carcinoma plasma has the potential of identifying candidate markers for early diagnosis and prognosis and/or to monitor disease recurrence. Here, we report on the utilization of a multi-dimensional fractionation approach (12P-M-LAC) and LC–MS/MS to comprehensively investigate clear cell renal cell carcinoma plasma collected before (disease) and after (non-disease) curative nephrectomy (n = 40). Proteins detected in the subproteomes were investigated via label-free quantification. Protein abundance analysis revealed a number of low-level proteins with significant differential expression levels in disease samples, including HSPG2, CD146, ECM...
TL;DR: The data support that the O-GlcNAc modification is involved in the regulation of adipocytokine secretion upon the induction of insulin resistance in human adipocytes and the classical method of hyperglycemia and hyperinsulinemia.
Abstract: Adipose tissue is both an energy storage depot and an endocrine organ. The impaired regulation of the secreted proteins of adipose tissue, known as adipocytokines, observed during obesity contributes to the onset of whole-body insulin resistance and the pathobiology of type 2 diabetes mellitus (T2DM). In addition, the global elevation of the intracellular glycosylation of proteins by O-linked β-N-acetylglucosamine (O-GlcNAc) via either genetic or pharmacological methods is sufficient to induce insulin resistance in both cultured cells and animal models. The elevation of global O-GlcNAc levels is associated with the altered expression of many adipocytokines. We have previously characterized the rodent adipocyte secretome during insulin sensitive and insulin resistant conditions. Here, we characterize and quantify the secretome and glycome of primary human adipocytes during insulin responsive and insulin resistant conditions generated by the classical method of hyperglycemia and hyperinsulinemia or by the pharmacological manipulation of O-GlcNAc levels. Using a proteomic approach, we identify 190 secreted proteins and report a total of 20 up-regulated and 6 down-regulated proteins that are detected in both insulin resistant conditions. Moreover, we apply glycomic techniques to examine (1) the sites of N-glycosylation on secreted proteins, (2) the structures of complex N- and O-glycans, and (3) the relative abundance of complex N- and O-glycans structures in insulin responsive and insulin resistant conditions. We identify 91 N-glycosylation sites derived from 51 secreted proteins, as well as 155 and 29 released N- and O-glycans respectively. We go on to quantify many of the N- and O-glycan structures between insulin responsive and insulin resistance conditions demonstrating no significant changes in complex glycosylation in the time frame for the induction of insulin resistance. Thus, our data support that the O-GlcNAc modification is involved in the regulation of adipocytokine secretion upon the induction of insulin resistance in human adipocytes.
TL;DR: The degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus is revealed.
Abstract: Cross-species viral transmission subjects parent and progeny alphaviruses to differential post-translational processing of viral envelope glycoproteins. Alphavirus biogenesis has been extensively studied, and the Semliki Forest virus E1 and E2 glycoproteins have been shown to exhibit differing degrees of processing of N-linked glycans. However the composition of these glycans, including that arising from different host cells, has not been determined. Here we determined the chemical composition of the glycans from the prototypic alphavirus, Semliki Forest virus, propagated in both arthropod and rodent cell lines, by using ion-mobility mass spectrometry and collision-induced dissociation analysis. We observe that both the membrane-proximal E1 fusion glycoprotein and the protruding E2 attachment glycoprotein display heterogeneous glycosylation that contains N-linked glycans exhibiting both limited and extensive processing. However, E1 contained predominantly highly processed glycans dependent on the host cell, with rodent and mosquito-derived E1 exhibiting complex-type and paucimannose-type glycosylation, respectively. In contrast, the protruding E2 attachment glycoprotein primarily contained conserved under-processed oligomannose-type structures when produced in both rodent and mosquito cell lines. It is likely that glycan processing of E2 is structurally restricted by steric-hindrance imposed by local viral protein structure. This contrasts E1, which presents glycans characteristic of the host cell and is accessible to enzymes. We integrated our findings with previous cryo-electron microscopy and crystallographic analyses to produce a detailed model of the glycosylated mature virion surface. Taken together, these data reveal the degree to which virally encoded protein structure and cellular processing enzymes shape the virion glycome during interspecies transmission of Semliki Forest virus.
TL;DR: Examination of the modulation of the total surface glycome of C. jejuni in response to temperature may help shed light on commensal and pathogenic mechanisms for this species.
TL;DR: The reviewed advances allow accurate modelling of carbohydrate-protein 3D co-complexes, but challenges remain in ranking the affinity of congeners.
TL;DR: The UUKV glycome is reported, revealing differential processing of the Gn and Gc virion glycoproteins, consistent with virion assembly in the medial Golgi apparatus, whereas oligomannose-type glycans required for DC-SIGN-dependent cellular attachment are predominant on Gc.
Abstract: Uukuniemi virus (UUKV) is a model system for investigating the genus Phlebovirus of the Bunyaviridae We report the UUKV glycome, revealing differential processing of the Gn and Gc virion glycoproteins Both glycoproteins display poly-N-acetyllactosamines, consistent with virion assembly in the medial Golgi apparatus, whereas oligomannose-type glycans required for DC-SIGN-dependent cellular attachment are predominant on Gc Local virion structure and the route of viral egress from the cell leave a functional imprint on the phleboviral glycome
TL;DR: Improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to include polysialylated N-glycans, and when combined together with the Glycomics Quintavariate Informed Quantification (GlyQ-IQ) recently described, this report is able to significantly extend glycan detection sensitivity and provide expanded glycan coverage.
Abstract: The N-glycan diversity of human serum glycoproteins, ie, the human blood serum N-glycome, is both complex and constrained by the range of glycan structures potentially synthesizable by human glycosylation enzymes The known glycome, however, has been further limited by methods of sample preparation, available analytical platforms, eg, based upon electrospray ionization-mass spectrometry (ESI-MS), and software tools for data analysis In this report several improvements have been implemented in sample preparation and analysis to extend ESI-MS glycan characterization and to include polysialylated N-glycans Sample preparation improvements included acidified, microwave-accelerated, PNGase F N-glycan release to promote lactonization, and sodium borohydride reduction, that were both optimized to improve quantitative yields and conserve the number of glycoforms detected Two-stage desalting (during solid phase extraction and on the analytical column) increased sensitivity by reducing analyte signal division
TL;DR: The results showed that the mono- and oligo-saccharide in Panax species were similar, including the glucan and pectin type of poly-Saccharides in different locations and different species of Panax, respectively, which may contribute to rational usage of polysaccharides fromPanax species.
Abstract: Multiple species of ginseng are well-known Chinese medicinal herbs. The glycome of Panax species has various beneficial effects; however, studies related to their systematic profiling are very limited. Therefore, the systematic profiling of the glycome of Panax species was investigated in this study. The sugars from different locations and different species of Panax (Panax ginseng, Panax quinquefolium, and Panax notoginseng) were prepared by microwave-assisted extraction. Free mono- and oligo-saccharides were identified by high-performance thin-layer chromatography (HPTLC). Furthermore, polysaccharides were compared and characterized by using saccharide mapping based on HPTLC analysis. The results showed that the mono- and oligo-saccharide in Panax species were similar, including the glucan and pectin type of polysaccharides in different locations and different species of Panax, respectively. The data are helpful to better understand the glycome of different species of Panax and may contribute to rational...
TL;DR: This Viewpoint highlights an emerging concept of cell surface engineering using synthetic nanoscale mimetics of native glycoconjugates to harness some of the unique biology of glycans, with an eye toward advancing stem cell-based neuroregenerative therapies.
Abstract: Glycans are important contributors to the development and function of the nervous system with enormous potential as therapeutic targets. However, a general lack of tools for tailoring the presentation of specific glycan structures on the surfaces of cells has left them largely unexplored in the biomedical context. In this Viewpoint, we briefly summarize the distinct challenges and complexities of the Glycome. We also highlight an emerging concept of cell surface engineering using synthetic nanoscale mimetics of native glycoconjugates to harness some of the unique biology of glycans, with an eye toward advancing stem cell-based neuroregenerative therapies.