Title : Carbohydrate composition and presence of a fucose-protein linkage in recombinant human
pro-urokinase
Abstract :
- A new post-translational modification site in the growth factor domain of urinary type plasminogen activator has been identified
- A glycopeptide containing the monosaccharide, fucose, covalently linked directly to the peptide backbone has been isolated from the tryptic digest of pro-urokinase expressed in a mouse hybridoma cell line Sp 2/0 Ag 14
- The glycopeptide was isolated by semi-preparative reversed phase high performance liquid chromatography
- The identity of a fucose containing peptide was confirmed by carbohydrate analysis, amino acid analysis and plasma desorption mass spectrometry (PDMS)
- A combination of these methodologies showed an equimolar ratio of peptide and fucose in the glycopeptide
- This modification is not detected without mass spectrometry because the fucose residue is hydrolyzed under standard acidic conditions of amino acid com position and N-terminal sequence analysis
- The site of attachment of fucose to the peptide has been localized towards the N-terminus (within first 23 amino acids) of the protein
- Also, the carbohydrate composition of recombinant pro-urokinase is reported
Output (sent_index, trigger,
protein,
sugar,
site):
- 0. composition, , -, pro-urokinase, pro
- 0. presence, , -, a fucose-protein linkage, pro
- 2. containing, , -, the monosaccharide, fucose,, glycopeptide
- 2. glycopeptide, , -, -, glycopeptide
- 3. glycopeptide, , -, -, glycopeptide
- 5. glycopeptide, , -, -, glycopeptide
- 7. attachment, , -, fucose, peptide
- 8. pro-urokinase, , -, the carbohydrate composition, pro
Output(Part-Of) (sent_index,
protein,
site):
*Output_Site_Fusion* (sent_index,
protein,
sugar,
site):