PXD002435

PXD002435 is an original dataset announced via ProteomeXchange.

Title	Glycosylation analysis of TNF-alpha-induced insulin resistance
Description	Glycoproteomics, TNF-alpha, membrane proteomics, insulin resistance, adipocyte, SILAC Insulin resistance (IR) is a complex process arising from both environmental and genetic perturbations and leads to a variety of diseases including type-2 diabetes (T2D). The expansion of adipose tissue during obesity results in secretion of proinflammatory cytokines which significantly impairs insulin sensitivity throughout the entire body. Inflammation modulates glycosylation in a variety of cell types however, an investigation of changes in glycosylation following inflammation-induced IR in adipocytes has not been performed. In the present study, we performed a quantitative N-glycomic analysis of TNF-alpha induced IR in adipocytes and identified the regulation of specific N-glycans including an increase in terminal di-galactose- and decrease in di-sialic acid-containing glycans with alpha-2,3 linkages. Quantitative analysis of the membrane-associated proteome in TNF-alpha treated adipocytes identified the regulation of specific glycosyltransferases and glycosidases which correlated with the regulated N-glycans. This included the up- and down-regulation of B4GalT5 and ST3Gal6, respectively at both the protein and mRNA level. To identify the protein and glycosylation sites modified with these regulated N-glycans, we performed a site-specific quantitative glycoproteomic analysis of enriched N-glycopeptides from TNF-alpha treated adipocytes with and without deglycosylation. The combined workflow provided a relative quantification of changes in protein abundance verses N-glycosylation occupancy verses site-specific N-glycans on a proteome-wide level. This revealed the modulation of a subset of N-glycan compositions on specific proteins including those previously associated with insulin-stimulated GLUT4 trafficking to the plasma membrane. Finally, knockdown of B4GalT5 with siRNA and analysis of released N-glycans resulted in the down-regulation of di-galactose containing glycans, confirming the involvement of this enzyme in the TNF-alpha regulated N-glycome.
HostingRepository	PRIDE
AnnounceDate	2015-11-09
AnnouncementXML	Submission_2015-11-09_03:49:00.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Benjamin Parker
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue; deaminated residue
Instrument	Orbitrap Fusion ETD; Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2015-06-29 01:38:56	ID requested
⏵ 1	2015-11-09 03:49:01	announced

Parker BL, Thaysen-Andersen M, Fazakerley DJ, Holliday M, Packer NH, James DE, Terminal Galactosylation and Sialylation Switching on Membrane Glycoproteins upon TNF-Alpha-Induced Insulin Resistance in Adipocytes. Mol Cell Proteomics, 15(1):141-53(2016) [pubmed]

curator keyword: Biomedical

submitter keyword: Glycoproteomics, TNF-alpha, membrane proteomics, insulin resistance, adipocyte, SILAC

Benjamin Parker
contact affiliation	The University of Sydney
contact email	benjamin.parker@sydney.edu.au
lab head
Benjamin Parker
contact affiliation	The University of Sydney
contact email	benjamin.parker@sydney.edu.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD002435
     1. Label: PRIDE project
     2. Name: Glycosylation analysis of TNF-alpha-induced insulin resistance