PMID: 14760718

 

    Legend: Gene, Sites

Title : Screening for N-glycosylated proteins by liquid chromatography mass spectrometry

Abstract :

1. In the last few years mass spectrometry has become the method of choice for characterization of post-translationally modified proteins
2. Whereas most protein chemical modifications are binary in the sense that only one change can be associated with a given residue, many different oligosaccharides can be attached to a glycosylation site residue
3. The detailed characterization of glycoproteins in complex biological samples is extremely challenging
4. However, information on N-glycosylation can be gained at an intermediary level
5. Here we demonstrate a procedure for mapping N-glycosylation sites in complex mixtures by reducing sample complexity and enriching glycoprotein content
6. Glycosylated proteins are selected by an initial lectin chromatography step and digested with endoproteinase Lys-C
7. Glycosylated peptides are then selected from the digest mixture by a second lectin chromatography step
8. The glycan components are removed with N-glycosidase F and the peptides digested with trypsin before analysis by on-line reversed-phase liquid chromatography mass spectrometry
9. Using two different lectins, concanavalin A and wheat germ agglutinin , this procedure was applied to human serum and a total of 86 N-glycosylation sites in 77 proteins were identified

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

• 0. N-glycosylated, , proteins, -, -
• 3. glycoproteins, , glycoproteins, -, -
• 5. N-glycosylation, , -, -, sites
• 5. glycoprotein, , glycoprotein, -, -
• 9. N-glycosylation, , -, -, sites

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

• 9. proteins, sites

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

 

Protein	NCBI ID	SENTENCE INDEX