PXD046194-1

PXD046194 is an original dataset announced via ProteomeXchange.

Title	Site-specific immobilization of the endosialidase for studying polysialic acid reveals QSOX2 is a novel polysialylated protein
Description	Polysialic acid (polySia) is a linear polymer of α2,8-linked sialic acid residues that is of fundamental biological interest due to its pivotal roles in the regulation of the nervous, immune, and reproductive systems in healthy human adults. PolySia is also dysregulated in several chronic diseases, including cancers and mental health disorders. However, the mechanisms underpinning polySia biology in health and disease remain largely unknown, in part due to the lack of tools with which to study the glycan. The polySia-specific hydrolase, endoneuraminidase NF (EndoN), and the catalytically inactive polySia lectin EndoNDM, have been extensively used for studying polySia. However, EndoN is heat stable and remains associated with cells after washing. When studying polySia in systems with multiple polysialylated species, the residual EndoN that cannot be removed confounds data interpretation. We developed a strategy for site-specific immobilization of EndoN and EndoNDM on streptavidin-coated magnetic or agarose beads. We showed that immobilizing EndoN improves enzyme usefulness by allowing for effective removal of the enzyme from samples, while retaining hydrolase activity. Additionally, immobilization of EndoNDM allowed for the enrichment of polysialylated proteins from complex mixtures for their identification via mass spectrometry. We identified QSOX2 as a novel polysialylated protein secreted from MCF-7 cells. This method of site-specific immobilization can be utilized for other enzymes and lectins to yield insight into glycobiology.
HostingRepository	PRIDE
AnnounceDate	2024-05-23
AnnouncementXML	Submission_2024-05-22_23:34:47.370.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Nichollas Scott
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	No PTMs are included in the dataset
Instrument	Orbitrap Fusion Lumos; Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2023-10-17 15:19:56	ID requested
⏵ 1	2024-05-22 23:34:48	announced

Hunter C, Derksen T, Makhsous S, Doll M, Perez SR, Scott NE, Willis LM, Site-specific immobilization of the endosialidase reveals QSOX2 is a novel polysialylated protein. Glycobiology, 34(5):(2024) [pubmed]

10.1093/glycob/cwae026;

submitter keyword: QSOX2, proteomics,EndoN, protein immobilization, biotinylation

Lisa M. Willis
contact affiliation	Department of Biological Sciences, University of Alberta, Edmonton, Canada Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, Canada
contact email	lisa.willis@ualberta.ca
lab head
Nichollas Scott
contact affiliation	University of Melbourne
contact email	nichollas.scott@unimelb.edu.au
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/2024/04/PXD046194

PRIDE project URI

• PRIDE
  1. PXD046194
     1. Label: PRIDE project
     2. Name: Site-specific immobilization of the endosialidase for studying polysialic acid reveals QSOX2 is a novel polysialylated protein