Dataset Information

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

ABSTRACT: 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

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive HF

ORGANISM(S): Homo Sapiens (human) Drosophila Melanogaster (fruit Fly)

SUBMITTER: Sergey Vakhrushev

LAB HEAD: Sergey Vakhrushev

PROVIDER: PXD024767 | Pride | 2021-04-07

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	200428_Lumos__LC1200__nanoFlex_Ieva_SARS_CoV2_h3_Chy_Chymo_90min_ETD_HCD_2ul.raw	Raw
	200428_Lumos__LC1200__nanoFlex_Ieva_SARS_CoV2_h3_T_G_GluC_Try_90min_ETD_HCD_2ul.raw	Raw
	200428_Lumos__LC1200__nanoFlex_Ieva_SARS_CoV2_i1_Chy_Chymo_90min_ETD_HCD_2ul.raw	Raw
	200428_Lumos__LC1200__nanoFlex_Ieva_SARS_CoV2_i1_T_G_GluC_Try_90min_ETD_HCD_2ul.raw	Raw
	200428_Lumos__LC1200__nanoFlex_Ieva_SARS_CoV2_i3_Chy_Chymo_90min_ETD_HCD_2ul.raw	Raw

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Publications

Site-Specific O-Glycosylation Analysis of SARS-CoV-2 Spike Protein Produced in Insect and Human Cells.

Bagdonaite Ieva I Thompson Andrew J AJ Wang Xiaoning X Søgaard Max M Fougeroux Cyrielle C Frank Martin M Diedrich Jolene K JK Yates John R JR Salanti Ali A Vakhrushev Sergey Y SY Paulson James C JC Wandall Hans H HH

Viruses 20210325 4

Enveloped viruses hijack not only the 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 location of O-glycans is controlled by a large family of initiating enzymes with variable expression in cells and tissues and hence ...[more]

PMID: 33806155

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