Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Mucin-type O-glycans enrich in white matter tracts

ABSTRACT: Protein O-glycosylation, the enzymatic attachment of carbohydrates to serine/threonine/tyrosine residues, is a large group of essential post-translational modifications involved in brain development and disease. However, studies of the most abundant O-glycan class in the mammalian brain, termed mucins or O-GalNAc glycans, are limited. Here, we determined the cellular specificity, spatial distribution, and protein carriers of O-GalNAc glycans in the mouse brain. Genetic inhibition of the O-GalNAc pathway in neurons or astrocytes had minimal effect on total O-GalNAc levels in the brain and failed to replicate the severe neurologic phenotype seen in rodent models and human congenital disorders of O-GalNAc synthesis. Further investigations revealed a surprising enrichment of O-GalNAc structures in white matter tracts and a spatial distribution distinct from other major carbohydrate structures in the brain including N-glycans. Using glycoproteomics, O-GalNAc glycans were identified on secretory proteins expressed in multiple cell types involved in brain development, synapse organization, and the extracellular matrix, including classical carriers of O-mannose glycans and proteoglycans. These findings highlight an overlooked aspect of O-GalNAc biology in white matter tracts of the brain and will inform future studies of development and disease.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Brain

SUBMITTER: Suttipong Suttapitugsakul

LAB HEAD: Robert G. Mealer

PROVIDER: PXD041631 | Pride | 2025-02-10

REPOSITORIES: pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	SS_MN_PNA_10282022_F1.raw	Raw
	SS_MN_PNA_10282022_F1.txt	Txt
	SS_MN_PNA_10282022_F10.raw	Raw
	SS_MN_PNA_10282022_F10.txt	Txt
	SS_MN_PNA_10282022_F2.raw	Raw

Items per page:

1 - 5 of 20

Similar Datasets

Large-scale and Site-specific Mapping of Intact O-glycans on Glycopeptides with IMPa

Project description:We employed the newly available IMPa O-glycoprotease from Pseudomonas aeruginosa for O-glycoproteomics analysis of cultured cells and tissues. The glycopeptides were extracted, purified, and conjugated to a solid support before an enzymatic cleavage by IMPa. O-glycopeptides were analyzed by EThcD, which allowed for a site-specific and global analysis of O-glycans, especially those sialylated and the Tn antigen. Through wild-type or SimpleCell HEK293 cells, we present two approaches, one for the analysis of total O-glycoproteome and the other for the Tn glycoproteome, and showed that IMPa and EThcD are necessary for confident localization of O-glycans on glycopeptides. We applied this method to study the O-glycoproteome of mouse brain, which revealed the high frequency of sialylated O-glycans and the Tn antigen on brain glycoproteins.

2023-08-28 | PXD037415 | Pride

Probing endogenous peptide acceptor substrate specificities of TbSTT3A and TbSTT3B

Project description:We performed glycoproteomics on endogenous parasite glycoproteins using sequential endoglycosidase-H and peptide-N-glycosidase-F digestions, the latter in the presence of H2[18O]. This allowed us to assess the relative occupancies of native N-glycosylation sites by endoglycosidase-H resistant N-glycans originating from Man5GlcNAc2-PP-dolichol transferred by TbSTT3A, and endoglycosidase-H sensitive N-glycans originating from Man9GlcNAc2-PP-dolichol transferred by TbSTT3B.

2017-09-20 | PXD007267 | Pride

Position-specific N- and O-glycosylation of the reactive centre loop impacts neutrophil elastase-mediated proteolysis of corticosteroid-binding globulin

Project description:Corticosteroid-binding globulin (CBG) delivers anti-inflammatory cortisol to inflamed tissues through the proteolytic cleavage of an exposed reactive centre loop (RCL) by neutrophil elastase (NE). We previously demonstrated that RCL-localised Asn347-linked N-glycans impact NE proteolysis, but a comprehensive structure-function characterisation of the RCL glycosylation is still required to better understand CBG glycobiology. Herein, we firstly performed RCL-centric glycoprofiling of serum-derived CBG to elucidate the Asn347-glycans and then used molecular dynamics (MD) simulations to study their impact on NE proteolysis. Importantly, we also identified novel O-glycosylation (di/sialyl T) across four RCL sites (Thr338/Thr342/Thr345/Ser350) of serum CBG close to the NE-targeted Val344-Thr345 cleavage site. A restricted N- and O-glycan co-occurrence pattern on the RCL involving exclusively Asn347 and Thr338 sites was experimentally observed and supported in silico by modelling of a CBG-GalNAc-transferase (GalNAc-T) complex with various RCL glycans. GalNAc-T2 and -T3 abundantly expressed by liver and gallbladder, respectively, showed in vitro a capacity to transfer GalNAc (Tn) to multiple RCL sites suggesting their involvement in RCL O-glycosylation. Recombinant CBG (rCBG) was then used to determine roles of RCL O-glycosylation through longitudinal NE-centric proteolysis experiments, which demonstrated that both sialo- (disialyl T) and asialo-glycans (T) decorating Thr345 inhibit NE proteolysis. Synthetic RCL O-glycopeptides expanded on these findings by showing that Thr345-Tn and Thr342-Tn confer strong and moderate protection against NE cleavage, respectively. MD substantiated that short Thr345-linked O-glycans abrogate NE interactions. In conclusion, we report on strategically-positioned and biologically-relevant CBG RCL glycans, which improve our understanding of mechanisms governing cortisol delivery to inflamed tissues.

2023-12-05 | PXD046405 | Pride

Electron-based dissociation is needed for O-glycopeptides derived from OpeRATOR proteolysis

Project description:The recently described O-glycoprotease OpeRATOR presents exciting opportunities for O-glycoproteomics. This bacterial enzyme purified from A. muciniphila cleaves N-terminally to serine and threonine residues that are modified with (preferably asialylated) O-glycans, providing orthogonal cleavage relative to canonical proteases (e.g., trypsin) to improve O-glycopeptide characterization with tandem mass spectrometry (MS/MS). O-glycopeptides with a modified N-terminal residue, such as those generated by OpeRATOR, present several potential benefits, perhaps the most notable being de facto O-glycosite localization without the need of glycan-retaining fragments in MS/MS spectra. Indeed, O-glycopeptides modified exclusively at the N-terminus would enable O-glycoproteomic methods to rely solely on collision-based fragmentation rather than electron-driven dissociation, but modified peptides would need to reliably contain only a single O-glycosite. Here we characterize the number of O-glycosites (i.e., missed cleavages) that are present in OpeRATOR-derived O-glycopeptides using methods that combine collision- and electron-based fragmentation. Our data show that over 50% of O-glycopeptides generated from combined digestion using OpeRATOR and trypsin contain multiple O-glycosites, indicating that collision-based fragmentation is not sufficient for OpeRATOR-centric studies and that electron-driven dissociation remains a requirement for site-specific O-glycopeptide characterization.

2020-11-03 | PXD020077 | Pride

Deconstruction of the GalNAc-Type O-Glycosylation Capacity of Human HEK293 Cells

Project description:Most proteins trafficking the secretory pathway of metazoan cells will acquire GalNAc-type O-glycosylation. GalNAc-type O-glycosylation is differentially regulated in cells by the expression of a repertoire of up to twenty genes encoding polypeptide GalNAc-transferase isoforms (GalNAc-Ts) that initiate O-glycosylation by catalyzing the attachment of GalNAc residues to select Ser and Thr residues. These GalNAc-Ts orchestrate the positions and patterns of O-glycans on proteins in poorly understood coordinated ways guided partly by the kinetic properties and substrate specificities of their catalytic domains and modulatory effects of their unique GalNAc-binding lectin domains. Here, we provide the hereto most comprehensive characterization of the non-redundant contributions of individual GalNAc-T isoforms to the O-glycoproteome of the human HEK293 cell line using quantitative differential O-glycoproteomics on a panel of isogenic HEK293 cell lines with knockout of GalNAc-T genes (GALNT1, T2, T3, T7, T10, or T11). We confirm that a major part of the O-glycoproteome is covered by redundancy, while distinct subsets of O-glycosites are covered by non-redundant GalNAc-T isoform-specific functions. We demonstrate that GalNAc-T7 and T10 as predicted from in vitro studies function in completion of high density O-glycosylated regions, while GalNAc-T11 selectively controls the site-specific O-glycosylation of low-density lipoprotein-related receptors in the linker regions between the ligand-binding LDLR class A repeats.

2019-06-24 | PXD009955 | Pride

Study of the differentially-expressed genes induced by the expession of two viral glycoproteins in muscle cells of adult zebrafish

Project description:A genomic approach was used to indentify the differences in the gene expression profiles of adult zebrafish injected with different viral glycoproteins. This study intended to study the mechanisms of immunogenicity of each glycoprotein (gpG of VHSV and SVCV) Fish were injected with plasmids encoding the different glycoproteins and muscle samples at the site of injection were compared to control samples at three days post-immunisation.

2013-08-08 | E-GEOD-49626 | biostudies-arrayexpress

O-GalNAc–glycosylated nuclear proteins

Project description:Biological functions of nuclear proteins are regulated by post-translational modifications (PTMs) that modulate gene expression and cellular physiology. However, the role of O-linked glycosylation (O-GalNAc) as a PTM of nuclear proteins in the human cell has not been previously reported. Here, we examined the initiation of O-GalNAc glycan biosynthesis, representing a novel PTM of nuclear proteins in the nucleus of human cells, with an emphasis on HeLa cells. Using affinity chromatography and MS analyses, we identified O-GalNAc glycosylated proteins in the nucleus and present solid evidence for O-GalNAc glycan synthesis in this organelle. The demonstration of O-GalNAc glycosylation of nuclear proteins in mammalian cells reported here has important implications for cell and chemical biology.

2019-04-17 | PXD013484 | Pride

mir-19 and mir-92 regulate lipid metabolism and energy balance

Project description:Profile gene expression change in 3T3 L1 adipocytes after overexpression of microRNA mir-17/18a, mir-18a/19a/20a, and mir-19b/92a

2015-06-19 | E-MTAB-2662 | biostudies-arrayexpress

O-glycan inhibitors generate aryl-glycans, induce apoptosis, and inhibit growth in colorectal cancer cell lines

Project description:Our studies provide direct evidence that O-glycosylation pathways play a role in the regulation of cell growth through apoptosis and proliferation pathways. Eight small molecular weight analogues of the GalNAc-alpha-1-O-serine/threonine structure based on 1-benzyl-2-acetamido-2- deoxy-alpha-O-D-galactopyranoside have been synthesised and tested in 5 human colorectal cancer cell lines. Three inhibitors, 1-benzyl-2-acetamido-2-deoxy-alpha-O-D-galactopyranoside and the corresponding 2-azido- and C-glycoside analogues, were screened in two colorectal cancer cell lines at 0.5mM and showed induction of apoptosis. Proliferation was down regulated in the same two cell lines with all three inhibitors, as detected by Ki67 staining and gene array. Treatment both cell lines with inhibitors led to changes in glycosylation detected with peanut lectin. The competitive action of the inhibitors resulted in the intracellular formation of 28 aryl-glycan products which were identified by MALDI and electrospray mass spectroscopy. The structures found map onto known O-glycosylation biosynthetic pathways and showed a differential pattern for each of the inhibitors in both cell lines. Gene array analysis of the glycogenes illustrated a pattern of glycosytransferases that matched the glycan structures found in glycoproteins and aryl-glycans formed in the PC/AA/C1/SB10C cells, however there was no action of the three inhibitors on glycogene transcript levels. The inhibitors act at both intermediary metabolic and genomic levels, resulting in altered protein glycosylation and arylglycan formation. These events may play a part in growth arrest. Keywords: Response to inhibitors, Apoptosis, Aryl-glycans, Benzyl-O-GalNAc, Growth Inhibition, O-Glycans

2008-08-18 | GSE12422 | GEO

Deconstruction of O-glycosylation â€“ Polypeptide GalNAc-T Isoforms Direct Distinct Subsets of the O-Glycoproteome

Project description:GalNAc-transferase (GalNAc-T) isoforms modify distinct subsets of the O-glycoproteome and GalNAc-type O-glycosylation is found on most proteins trafficking through the secretory pathway in metazoan cells. The O-glycoproteome is regulated by up to 20 polypeptide GalNAc-Ts and the contributions and biological functions of individual GalNAc-Ts are poorly understood. Here, we used a zinc-finger nuclease (ZFN)-directed knockout strategy to probe the contributions of the major GalNAc-Ts (GalNAc-T1 and T2) in liver cells, and explore how the GalNAc-T repertoire quantitatively affects the O-glycoproteome. We demonstrate that the majority of the O-glycoproteome is covered by redundancy, whereas distinct subsets of substrates are modified by non-redundant functions of GalNAc-T1 and T2. Differential transcriptomic analysis indicates that loss of function of a GalNAc-T induces specific transcriptional response. The non-redundant O-glycoproteome subsets for and the transcriptional responses for each isoform appeared to be related to different cellular processes, and for the GalNAc-T2 isoform supporting a role in lipid metabolism. The results demonstrate that GalNAc-Ts have non-redundant glycosylation functions, and that these may affect distinct cellular processes. The data provides a comprehensive resource for unique substrates for individual GalNAc-Ts. Our study provides a new view on the regulation of the O-glycoproteome, suggesting that the plurality of GalNAc-Ts arose to regulate distinct protein functions and cellular processes.

2015-09-30 | E-MTAB-3844 | biostudies-arrayexpress

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data