Quantitative N-linked Glycoproteomics of Myocardial Ischemia and Reperfusion Injury Reveals Early Remodeling in the Extracellular Environment

Abstract

changes. These were mainly from predicted extracellular matrix and basement membrane proteins that are implicated in cardiac remodeling. Analysis of N-glycans released from myocardial proteins suggest that the observed changes were not due to significant alterations in N-glycan structures. Altered proteins included the collagen-laminin-integrin complexes and collagen assembly enzymes, cadherins, mast cell proteases, proliferation-associated secreted protein acidic and rich in cysteine, and microfibril-associated proteins. The data suggest that cardiac remodeling is initiated earlier during reperfusion than previously hypothesized.

Hypoxia-induced Changes to Integrin α 3 Glycosylation Facilitate Invasion in Epidermoid Carcinoma Cell Line A431

“…In 2011, Parker et al enriched and quantified iTRAQ-labeled N-glycopeptides during myocardial ischemia and reperfusion injury via sequential enrichment with TiO 2 followed by IP-ZIC-HILIC (Zwitterionic hydrophilic interaction chromatography with ion-pairing) for the unbound fraction. More than 400 N-linked glycopeptides were statistically quantified, and 80 glycosylation sites were found altered, in which the glycosylation of several integrin subunits (ITGA6/7/V and ITGB1) was also found up-regulated following ischemia and reperfusion injury (59). These results were consistent with our findings.…”

A Novel Method for the Simultaneous Enrichment, Identification, and Quantification of Phosphopeptides and Sialylated Glycopeptides Applied to a Temporal Profile of Mouse Brain Development

“…After incubation, the peptide mixture was diluted 1:1 with 1% TFA and purified on an Oligo R3 reversed phase micro-column as described previously (17) and the eluted peptides were dried by vacuum centrifugation. An aliquot of the enriched phospho-and N-linked sialylated glyco-peptides without PNGase F treatment was analyzed directly by nano-LC-MS/MS to identify chemically deamidated peptides occurring within the sequon for N-glycosylation (NxS/T/C) that could be interpreted as false positives after PNGase F treatment (21,22).…”

“…We have previously demonstrated that false positive glycosylations may occur in large-scale datasets by mis-annotation of chemically deamidated Asn residues within an N-linked sequon (21,22). To control for this, we employed the method set out in (21). Analysis of an aliquot of the enriched phospho-and N-linked sialylated glycopeptides was undertaken without PNGase F treatment and interrogation of the TiO 2 flow-through resulted in the identification of 202 unique deamidated peptides with 39 containing a deamidation within the motif for N-linked glycosylation (NxS/T/C).…”

“…We have previously demonstrated that false positive glycosylations may occur in large-scale datasets by mis-annotation of chemically deamidated Asn residues within an N-linked sequon (21,22). To control for this, we employed the method set out in (21).…”

Large‐scale N‐glycoproteome map of rat brain tissue: Simultaneous characterization of insoluble and soluble protein fractions

“…Furthermore, the rest 415 sites, which were not referred in the Swiss-Prot database, were identified in this work (Fig. 2); about 36% of them (148 sites) were found in a very recent study by Parker et al [28], and the other 267 sites, to the best of our knowledge, were hitherto not reported in rat. The detailed information of the identified N-glycosites was listed in Supporting Information Table 1, sheet 2.…”