PXD049172

PXD049172 is an original dataset announced via ProteomeXchange.

Title	Glycoproteomic and single-protein glycomic analyses reveal zwitterionic N-glycans on natural and recombinant proteins derived from insect cells
Description	Insect cells are a convenient cell factory to produce recombinant glycoproteins. Their glycosylation potential is believed to be simple, needing primarily addition of glycosyltransferases to humanize the recombinant products. In this study, the native glycoproteome of Spodoptera frugiperda Sf9 and Trichoplusia ni High Five cells examined using an LC-MS/MS approach revealed not only which proteins are N-glycosylated, but that the reconstructed N-glycomes contain glucuronylated and phosphorylcholine-modified glycans in addition to the typical oligomannosidic and fucosylated structures, These data were corroborated by a parallel MALDI-TOF MS/MS analysis of N-glycosidase-released oligosaccharides. Molecular modelling analysis of one endogenous Sf9 glycoprotein correlated the occurrence of complex and oligomannosidic N-glycans with the accessibility of the occupied N-glycosylation sites. Further, we showed that the N-glycans of influenza haemagglutinins and SARS-CoV-2 spike glycoprotein produced in Spodoptera cells possess a number of glycan structures modified with phosphorylcholine, but core difucosylation was minimal; in contrast, the Trichoplusia-produced haemagglutinin had only traces of the former type, while the latter were dominant. Detection of phosphorylcholine on these glycoproteins correlated with binding to human C-reactive protein. In conclusion, not just oligomannosidic or truncated paucimannosidic N-glycans, but structures with non-human features occur on both natural and recombinant glycoproteins derived from insect cell lines.
HostingRepository	PRIDE
AnnounceDate	2025-05-06
AnnouncementXML	Submission_2025-05-06_00:42:34.945.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Johannes Stadlmann
SpeciesList	scientific name: Spodoptera frugiperda; NCBI TaxID: 7108; scientific name: Trichoplusia ni; NCBI TaxID: 7111;
ModificationList	monohydroxylated residue; complex glycosylation; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2024-02-05 03:02:23	ID requested
⏵ 1	2025-05-06 00:42:35	announced

10.1016/J.MCPRO.2025.100981;

submitter keyword: phosphorylcholine, fucose,Baculovirus expression, N-glycans, insect cells

Johannes Stadlmann
contact affiliation	University of Natural Resources and Life Sciences (BOKU), Muthgasse 18, 1190 Vienna, Austria.
contact email	j.stadlmann@boku.ac.at
lab head
Johannes Stadlmann
contact affiliation	University of Natural Resources and Life Sciences (BOKU), Muthgasse 18, 1190 Vienna, Austria.
contact email	j.stadlmann@boku.ac.at
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD049172
     1. Label: PRIDE project
     2. Name: Glycoproteomic and single-protein glycomic analyses reveal zwitterionic N-glycans on natural and recombinant proteins derived from insect cells