PMID: 15253437

 

    Legend: Gene, Sites

Title : A new strategy for identification of N-glycosylated proteins and unambiguous assignment of their glycosylation sites using HILIC enrichment and partial deglycosylation

Abstract :

1. Characterization of glycoproteins using mass spectrometry ranges from determination of carbohydrate-protein linkages to the full characterization of all glycan structures attached to each glycosylation site
2. In a novel approach to identify N-glycosylation sites in complex biological samples, we performed an enrichment of glycosylated peptides through hydrophilic interaction liquid chromatography (HILIC) followed by partial deglycosylation using a combination of endo-beta-N-acetylglucosaminidases (EC 3.2.1.96)
3. After hydrolysis with these enzymes , a single N-acetylglucosamine (GlcNAc) residue remains linked to the asparagine residue
4. The removal of the major part of the glycan simplifies the MS/MS fragment ion spectra of glycopeptides , while the remaining GlcNAc residue enables unambiguous assignment of the glycosylation site together with the amino acid sequence
5. We first tested our approach on a mixture of known glycoproteins , and subsequently the method was applied to samples of human plasma obtained by lectin chromatography followed by 1D gel-electrophoresis for determination of 62 glycosylation sites in 37 glycoproteins

Output (sent_index, trigger, protein, sugar, site):

• 0. N-glycosylated, , proteins, -, -
• 0. glycosylation, , -, -, sites
• 1. attached, , -, all glycan structures, site
• 1. glycoproteins, , glycoproteins, -, -
• 1. glycosylation, , -, -, site
• 2. N-glycosylation, , -, -, sites
• 2. glycosylated, , -, -, peptides
• 4. glycopeptides, , -, -, glycopeptides
• 4. glycosylation, , -, -, site
• 5. glycoproteins, , glycoproteins, -, -
• 5. glycosylation, , -, -, sites

Output(Part-Of) (sent_index, protein, site):

• 5. glycoproteins, sites

*Output_Site_Fusion* (sent_index, protein, sugar, site):

 

Protein	NCBI ID	SENTENCE INDEX