Dataset Information

Site-specific glycosylation of donkey milk lactoferrin investigated by High Resolution Mass Spectrometry

ABSTRACT: A comprehensive glycosylation profile of donkey lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO2 and HILIC enrichment, reversed-phase high performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. The results obtained allowed the identification of 26 different glycan structures, including high mannose, complex and hybrid N-glycans, linked to the protein backbone via an amide bond to asparagine residues located at the positions 137, 281 and 476. Altogether, the N-glycan structures determined revealed that in donkey milk lactoferrin most of the N-glycans identified are neutral complex/hybrid. Actually, 10 neutral non-fucosylated complex/hybrid N-glycans and 4 neutral fucosylated complex/hybrid N-glycans were found. In addition, 2 high mannose N-glycans, 4 sialylated fucosylated complex/hybrid N-glycans and 6 sialylated non-fucosylatedN-glycans, one of which containing N-glycolylneuramin acid (Neu5Gc), were found. A comparison of the glycosylation profile of donkey milk lactoferrin with respect to that of human, bovine and goat milk lactoferrin is reported.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Equus Asinus (donkey)

TISSUE(S): Milk

SUBMITTER: Salvatore Foti

LAB HEAD: Salvatore Foti

PROVIDER: PXD004289 | Pride | 2016-08-25

REPOSITORIES: Pride

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Publications

Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry.

Gallina Serafina S Saletti Rosaria R Cunsolo Vincenzo V Muccilli Vera V Foti Salvatore S Roepstorff Peter P Rasmussen Morten I MI

Amino acids 20160822 12

A comprehensive monosaccharide composition of the N-glycans of donkey milk lactoferrin, isolated by ion exchange chromatography from an individual milk sample, was obtained by means of chymotryptic digestion, TiO<sub>2</sub> and HILIC enrichment, reversed-phase high-performance liquid chromatography, electrospray mass spectrometry, and high collision dissociation fragmentation. The results obtained allowed identifying 26 different glycan structures, including high mannose, complex and hybrid N-g ...[more]

PMID: 27550041

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Project description:Glycobiology plays a central role in prostate cancer (PCa), as evidenced by the aberrant glycosylation in PCa-derived glycoproteins. Understanding the protein glycosylation changes underling PCa onset and progression may open opportunities for the discovery of novel biomarkers for PCa detection and stratification as well as targets for PCa metastasis. Advances in mass spectrometry-based glycomics and glycoproteomics have opened up exciting avenues for deep characterisation of the immensely, yet poorly understood complex glycoproteome. In this study, we have used integrated glycomics and glycoproteomics to explore the protein, cell and tumour-grade specific N- and O- glycosylation signatures in surgically-removed fresh PCa tissues grouped into five PCa disease grades and tissues from benign prostatic hyperplasia patients (BPH). Porous graphitised carbon–liquid chromatography (PGC–LC)-MS/MS analysis was used to profile the N-and O-glycome, revealing PCa grade specific N and O glycosylation changes, including oligomannosidic and paucimannosidic, bisecting-GlcNAc and highly branched tri- and tetra-antennary fucosylated and sialylated N-glycans as well as sialylated core 1 and 2 type O-glycans. High resolution LC-MS/MS was then employed to capture the micro and macroheterogeneity of 1,085 N-glycosites (>7,400 unique N-glycopeptides) from 540 N-glycoproteins and 360 O-glycosites (>500 unique O-glycopeptides) from 178 O-glycoproteins and reveal protein and cell specific N- and O- glycosylation changes associated with PCa progression. We also showed PCa grade-specific increase in the expression of oligosaccharyltransferase (OST) subunits that correlate with changes in the site occupancy of N-glycoproteins. In conclusion, this study shows that integrated glycomics and glycoproteomics can provide unprecedented insight into key molecular features and site-specific glycan heterogeneity contributing to the highly diverse tumour-microenvironment and allow us to deconstruct the regulation of the protein, cell and tumour-grade-specific glycosylation associated with PCa onset and development.

Dataset Information

Site-specific glycosylation of donkey milk lactoferrin investigated by High Resolution Mass Spectrometry

Publications

Site-specific glycosylation of donkey milk lactoferrin investigated by high-resolution mass spectrometry.

Similar Datasets

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