PXD027605-1

PXD027605 is an original dataset announced via ProteomeXchange.

Title	Structural O-Glycoform Heterogeneity of the SARS-CoV-2 Spike Protein Receptor-Binding Domain Revealed by Top-Down Mass Spectrometry
Description	Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) utilizes an extensively glycosylated surface spike (S) protein to mediate host cell entry and the S protein glycosylation plays key roles in altering viral binding/function and infectivity. However, the molecular structures and glycan heterogeneity of the new O-glycans found on the S protein regional-binding domain (S-RBD) remain cryptic because of the challenges in intact glycoform analysis by conventional bottom-up glycoproteomic approaches. Here, we report the complete structural elucidation of intact O-glycan proteoforms through a hybrid native and denaturing top-down mass spectrometry (MS) approach employing both trapped ion mobility spectrometry (TIMS) quadrupole time-of-flight and ultrahigh-resolution Fourier transform ion cyclotron resonance (FTICR)-MS. Native top-down TIMS-MS/MS separates the protein conformers of the S-RBD to reveal their gas-phase structural heterogeneity, and top-down FTICR-MS/MS provides in-depth glycoform analysis for unambiguous identification of the glycan structures and their glycosites. A total of eight O-glycoforms and their relative molecular abundance are structurally elucidated for the first time. These findings demonstrate that this hybrid top-down MS approach can provide a high-resolution proteoform-resolved mapping of diverse O-glycoforms of the S glycoprotein, which lays a strong molecular foundation to uncover the functional roles of their O-glycans. This proteoform-resolved approach can be applied to reveal the structural O-glycoform heterogeneity of emergent SARS-CoV-2 S-RBD variants, as well as other O-glycoproteins in general.
HostingRepository	MassIVE
AnnounceDate	2021-07-27
AnnouncementXML	Submission_2021-07-27_16:48:50.383.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	David Roberts
SpeciesList	scientific name: Homo sapiens; common name: human; NCBI TaxID: 9606;
ModificationList	unknown modification; unknown modification; unknown modification; Acetyl; sulfated residue; unknown modification; unknown modification; unknown modification
Instrument	solariX; timsTOF Pro

Revision	Datetime	Status	ChangeLog Entry
0	2021-07-27 13:00:19	ID requested
⏵ 1	2021-07-27 16:48:50	announced

no publication

submitter keyword: O-Glycoform, O-glycosylation, COVID-19, SARS-CoV-2, Spike protein, regional binding domain , top-down mass spectrometry

Ying Ge
contact affiliation	University of Wisconsin-Madison
contact email	ying.ge@wisc.edu
lab head
David S. Roberts
contact affiliation	University of Wisconsin-Madison
contact email	dsroberts@wisc.edu
lab head
David Roberts
contact affiliation	University of Wisconsin-Madison
contact email	dsroberts@wisc.edu
dataset submitter

MassIVE dataset URI

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