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):

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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Project description:Background: Gene expression variation is a phenotypic trait of particular interest as it represents the initial link between genotype and other phenotypes. Analyzing how such variation apportions among and within groups allows for the evaluation of how genetic and environmental factors influence such traits. It also provides opportunities to identify genes and pathways that may have been influenced by non-neutral processes. Here we use a population genetics framework and next generation sequencing to evaluate how gene expression variation is apportioned among four human groups in a natural biological tissue, the placenta. Results: We estimate that on average, 33.2%, 58.9% and 7.8% of the placental transcriptome is explained by variation within individuals, among individuals and among human groups, respectively. Additionally, when technical and biological traits are included in models of gene expression they account for roughly 2% of total gene expression variation. Notably, the variation that is significantly different among groups is enriched in biological pathways associated with immune response, cell signaling and metabolism. Many biological traits demonstrated correlated changes in expression in numerous pathways of potential interest to clinicians and evolutionary biologists. Finally, we estimate that the majority of the human placental transcriptome (65% of expressed genes) exhibits expression profiles consistent with neutrality; the remainder are consistent with stabilizing selection (26%), directional selection (4.9%), or diversifying selection (4.8%). Conclusion: We apportion placental gene expression variation into individual, population and biological trait factors and identify how each influence the transcriptome. Additionally, we advance methods to associate expression profiles with different forms of selection. Placental mRNA was sequenced on an Illumina GAIIx. Samples were derived from 4 human groups, 10 individuals per group, 2 samples per individual