LC-MS/MS characterization of O-glycosylation sites and glycan structures of human cerebrospinal fluid glycoproteins.
Article Details
-
Citation
Copy to clipboard -
Halim A, Ruetschi U, Larson G, Nilsson J
LC-MS/MS characterization of O-glycosylation sites and glycan structures of human cerebrospinal fluid glycoproteins.
J Proteome Res. 2013 Feb 1;12(2):573-84. doi: 10.1021/pr300963h. Epub 2013 Jan 11.
- PubMed ID
-
23234360 [View in PubMed]
- Abstract
-
The GalNAc O-glycosylation on Ser/Thr residues of extracellular proteins has not been well characterized from a proteomics perspective. We previously reported a sialic acid capture-and-release protocol to enrich tryptic N- and O-glycopeptides from human cerebrospinal fluid glycoproteins using nano-LC-ESI-MS/MS with collision-induced dissociation (CID) for glycopeptide characterization. Here, we have introduced peptide N-glycosidase F (PNGase F) pretreatment of CSF samples to remove the N-glycans facilitating the selective characterization of O-glycopeptides and enabling the use of an automated CID-MS(2)/MS(3) search protocol for glycopeptide identification. We used electron-capture and -transfer dissociation (ECD/ETD) to pinpoint the glycosylation site(s) of the glycopeptides, identified as predominantly core-1-like HexHexNAc-O- structure attached to one to four Ser/Thr residues. We characterized 106 O-glycosylations and found Pro residues preferentially in the n - 1, n + 1, and/or n + 3 positions in relation to the Ser/Thr attachment site (n). The characterization of glycans and glycosylation sites in glycoproteins from human clinical samples provides a basis for future studies addressing the biological and diagnostic importance of specific protein glycosylations in relation to human disease.
DrugBank Data that Cites this Article
- Polypeptides
-
Name UniProt ID Neuron-specific vesicular protein calcyon Q9NYX4 Details Apolipoprotein A-II P02652 Details Cathepsin D P07339 Details Apolipoprotein E P02649 Details Neuroendocrine convertase 1 P29120 Details Plasma protease C1 inhibitor P05155 Details Neurocan core protein O14594 Details Complement C4-A P0C0L4 Details Apolipoprotein C-II P02655 Details Nucleobindin-1 Q02818 Details Alpha-1-antichymotrypsin P01011 Details Amyloid-like protein 1 P51693 Details Prostaglandin-H2 D-isomerase P41222 Details Hemopexin P02790 Details