Dataset Information

O-GlcNAcome analysis in human placenta

ABSTRACT: In this study, we expand on this major advancement in O-GlcNAc enrichment protocols by adapting this approach to human-banked samples, which often presents their significant challenges. Indeed, patient samples are often banked without proper washing, thus contaminating cells with abundant O-glycosylated blood protein. These glycoproteins, such as human serum albumin, antibodies, and hemoglobin, pollute MS acquisition, masking the less abundant intracellular proteins and, even more, their O-GlcNAcylated forms. Thus, using freshly banked placentas, we present an adapted method to prepare human placental samples with naturally high blood content (13). We then enriched for O-GlcNAcylated protein using the PTMscan antibodies previously mentioned (12). However, our adapted protocol yielded over 60% of HexNAc peptide on total peptides. Per sample, we obtained up to 2,600 HexNAc-containing PSMs, 1,090 unique HexNAc peptides, and more than 1,000 unique HexNAc proteins using 45mg of starting placenta. Furthermore, we present a bioinformatic workflow to sort these HexNAc PSMs and extract the most confident O-GlcNAcylated proteins and sites. Thus, we confidentially identified 641 O-GlcNAcylated proteins, including up to 146 new proteins and 730 new sites within the 2 placental samples analyzed. Furthermore, we enriched for over 50 placenta-enriched O-GlcNAcylated proteins, such as RPA4, representing a new biomarker of the placental O-GlcNAcylation.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Syncytiotrophoblast Cell, Placenta

SUBMITTER: Stephanie Olivier-Van Stichelen

LAB HEAD: Stephanie Olivier-Van Stichelen

PROVIDER: PXD047766 | Pride | 2024-02-14

REPOSITORIES: Pride

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			Action	DRS
	2164-2178_Porter_221118-230309_all-combined_01.mzML	Mzml
	2164-2178_Porter_221118-230309_all-combined_01.mzid.gz	Mzid
	2164_Porter_221118_Ts024_A-180_01.raw	Raw
	2164_Porter_221118_Ts024_A-180_02.raw	Raw
	2164_Porter_221118_Ts024_A-90_01.raw	Raw

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Publications

Studying the O-GlcNAcome of human placentas using banked tissue samples.

Luna Sarai S Malard Florian F Pereckas Michaela M Aoki Mayumi M Aoki Kazuhiro K Olivier-Van Stichelen Stephanie S

Glycobiology 20240401 4

O-GlcNAcylation is a dynamic modulator of signaling pathways, equal in magnitude to the widely studied phosphorylation. With the rapid development of tools for its detection at the single protein level, the O-GlcNAc modification rapidly emerged as a novel diagnostic and therapeutic target in human diseases. Yet, mapping the human O-GlcNAcome in various tissues is essential for generating relevant biomarkers. In this study, we used human banked tissue as a sample source to identify O-GlcNAcylated ...[more]

PMID: 38253038

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