PXD044353-1

PXD044353 is an original dataset announced via ProteomeXchange.

Title	Genotype-dependent N-glycosylation and newly discovered O-glycosylation affecting HRG plasmin cleavage
Description	Histidine-rich glycoprotein (HRG) is an abundant plasma glycoprotein with three reported N-glycosylation sites, which integrates many biological processes such as antiangiogenic activity, immune complex clearance and pathogen clearance. Importantly, the protein is known to have five genetic variants with minor allele frequencies of more than 10%, meaning these exist with substantial frequency in the human population. Among them, Pro204Ser can induce a new N-glycosylation site at Asn202. Considerable research has been performed into the biological activity of HRG, while research on its glycosylation is rare. To close this knowledge gap, we used C18-based LC-MS/MS to investigate the glycosylation characteristics of HRG from human plasma, recombinant Chinese hamster ovary (CHO) cell lines and recombinant human embryonic kidney (HEK293) cell lines with targeted mutations. Within human plasma endogenous HRG, every N-glycosylation site proved dominant with a sialylated diantennary complex-type glycan. For the recombinant HRGs, on the other hand, glycans showed different antennarities, sialylation and core-fucosylation, as well as the appearance of oligomannose glycans, LacdiNAc and antennary fucosylation. Furthermore, we discovered a previously unreported O-glycosylation site on residues Thr273/Thr274, which showed an approximate 90% glycan occupancy in all HRG types. To investigate the relevance of HRG glycosylation characteristics and its biological function, we set up an assay to study the plasmin cleavage of HRG under various conditions. In doing so, we showed that the sialylation of the new O-glycan, as well as the genotype-dependent N-glycosylation, significantly influenced the plasmin cleavage of HRG.
HostingRepository	PRIDE
AnnounceDate	2024-05-21
AnnouncementXML	Submission_2024-05-21_12:07:20.282.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Yang Zou
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	carboxylated residue; phosphorylated residue; monohydroxylated residue
Instrument	Orbitrap Fusion Lumos; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2023-08-04 05:20:05	ID requested
⏵ 1	2024-05-21 12:07:21	announced

10.1016/j.jbc.2024.105683;

Zou Y, Pronker MF, Damen JMA, Heck AJR, Reiding KR, Genotype-dependent N-glycosylation and newly exposed O-glycosylation affect plasmin-induced cleavage of histidine-rich glycoprotein (HRG). J Biol Chem, 300(3):105683(2024) [pubmed]

submitter keyword: histidine-rich glycoprotein (HRG), O-glycosylation, glycoproteomics, plasmin cleavage,N-glycosylation

Karli Reiding
contact affiliation	1Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Padualaan 8, 3584 CH, Utrecht, The Netherlands 2Netherlands Proteomics Center, Padualaan 8, 3584 CH, Utrecht, The Netherlands
contact email	k.r.reiding@uu.nl
lab head
Yang Zou
contact affiliation	Utrecht University
contact email	y.zou2@uu.nl
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/05/PXD044353

PRIDE project URI

• PRIDE
  1. PXD044353
     1. Label: PRIDE project
     2. Name: Genotype-dependent N-glycosylation and newly discovered O-glycosylation affecting HRG plasmin cleavage