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Characterisation of the human hemostatic O-glycoproteome

ABSTRACT: The hemostatic system comprises platelet aggregation, coagulation and fibrinolysis, and is critical to the maintenance of vascular integrity. Multiple studies indicate that glycans play an important role in maintaining the hemostatic system, however, most investigations have focused on N-glycans and little is known about the location and function of O-glycans. Here we performed the first systematic analysis of hemostatic O-glycosylation using lectin affinity chromatography coupled to LC-MS/MS to determine the precise location of O-glycans in human plasma, platelets, and endothelial cells. We identified the hitherto largest O-glycoproteome from native tissue, demonstrating that O-glycosylation is a ubiquitous modification of hemostatic proteins.

INSTRUMENT(S): LTQ Orbitrap Velos, Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Umbilical Vein Endothelial Cell, Blood Platelet, Platelet, Blood Plasma, Endothelial Cell

SUBMITTER: Sarah King

LAB HEAD: Hans Wandall

PROVIDER: PXD004590 | Pride | 2017-03-15

REPOSITORIES: Pride

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CHO O-Mannosylation independent of POMT1/2

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2017-05-18 | PXD004341 | Pride

Human O-Mannosylation independent of POMT1/2

2017-05-18 | PXD004358 | Pride

1. Site-specific O-glycosylation analysis of SARS-CoV-2 spike protein produced in insect and human cells

Project description:2. Enveloped viruses hijack not only host translation processes, but also its glycosylation machinery and to a variable extent cover viral surface proteins with tolerogenic host-like structures. SARS-CoV-2 surface protein S presents as a trimer on the viral surface and is covered by a dense shield of N-linked glycans, and a few O-glycosites have been reported. The biosynthetic nature of O-glycosylation is highly variable and is affected by initiating enzyme repertoires in different tissues and cell types, and therefore warrants a more thorough investigation. Here, we used our well-established O-glycoproteomic workflows to map the precise positions of O-linked glycosylation sites on three different entities of protein S – insect cell or human cell-produced ectodomains, or insect cell-derived receptor binding domain (RBD). In total 25 O-glycosites were identified, with similar patterns in the two ectodomains of different cell origin, and a distinct pattern of the monomeric RBD. Strikingly, 16 out of 25 O-glycosites were located within three amino acids from known N-glycosites. However, O-glycosylation was primarily found on peptides that were unoccupied by N-glycans, and otherwise had low overall occupancy. This suggests possible complimentary functions of O-glycans in immune shielding and negligible effects of O-glycosylation on subunit vaccine design for SARS-CoV-2

2021-04-07 | PXD024767 | Pride

Structure and dynamics of the Arabidopsis O-fucosyltransferase SPINDLY

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O-Fucosylation of CPN20 by SPINDLY Derepresses Abscisic Acid Signaling during Seed Germination and Seedling Development--Partial Submission

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