PXD059329

PXD059329 is an original dataset announced via ProteomeXchange.

Title	Endoglycosidases- treated N-glycosylation site identification
Description	Glycoproteins, representing over 50% of human proteins and most biopharmaceuticals, are crucial for regulating various biological processes. The complexity of multiple glycosylation sites often leads to incomplete sequence coverage and ambiguous glycan modification profiles. Here, we developed an integrative mass spectrometry-based approach for decoding unknown glycoproteins, which is featured with the combination of deglycosylation-mediated de novo sequencing with glycosylation site characterization. We utilized enzymatic deglycosylation of N-/ O- glycans to achieve comprehensive sequence coverage. Additionally, EThcD fragmentation enables the identification of high-quality long peptides, facilitating precise protein assembly. We subsequently applied this method to de novo sequencing of the highly glycosylated therapeutic fusion protein Etanercept (Enbrel®). We also sequenced three new tumor necrosis factor receptor (TNFR): Fc-fusion biologics with largely unknown sequences, unveiling subtle distinctions in the primary sequences. Furthermore, we characterized N- and O-glycosylation modifications of these proteins at subunit, glycopeptide, and glycan levels. This strategy bridges the gap between the de novo sequencing and glycosylation modification, providing comprehensive information of the primary structure and glycosylation modifications for glycoproteins. Notably, our method could be a robust solution for accurate sequencing of the glycoproteins and has practical value not only in basic research but also in the biopharmaceutical industry.
HostingRepository	iProX
AnnounceDate	2024-12-30
AnnouncementXML	Submission_2024-12-31_01:02:20.585.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Jing Gao
SpeciesList	scientific name: Homo sapiens; NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2024-12-31 01:01:19	ID requested
⏵ 1	2024-12-31 01:02:20	announced

Dataset with its publication pending

submitter keyword: de novo sequencing, N-glycosylation, Etanercept, tumor necrosis factor receptor (TNFR): Fc-fusion biologics

Zhou Hu
contact affiliation	Shanghai Institute of Materia Medica, Chinese Academy of Sciences
contact email	zhouhu@simm.ac.cn
lab head
Jing Gao
contact affiliation	Shanghai Institute of Materia Medica, Chinese Academy of Sciences
contact email	jinggao@simm.ac.cn
dataset submitter

iProX dataset URI