Characterization of the primary structure of TNK-tissue plasminogen activator using LC-MS / 药学学报
Acta Pharmaceutica Sinica
; (12): 896-900, 2013.
Article
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-259533
Responsible library:
WPRO
ABSTRACT
The primary structure of TNK-tissue plasminogen activator (TNK-tPA) was characterized using liquid chromatography-mass spectrometry (LC-MS). Firstly, the molecular mass of deglycosylated protein was measured. Then peptide mass mapping and MS/MS of the reduced, alkylated and trypsin-digested sample were tested and analyzed so as to verify its amino acid sequence and identify post-translational modifications. Results show that the amino acid sequence was consistent with designed structure; about 5% of M207 was oxidized; T61 was fucosylated with -80% occupancy; N103, N448 and N184 (-15% occupancy) were glycosylated with complex-type oligosaccharides. LC-MS coupled with proper sample pretreatment is approved to be a rapid and powerful approach to characterize the primary structure of TNK-tPA.
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Mass Spectrometry
/
Glycosylation
/
Chemistry
/
Protein Processing, Post-Translational
/
Chromatography, Liquid
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Amino Acid Sequence
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Tissue Plasminogen Activator
/
Molecular Weight
Language:
Chinese
Journal:
Acta Pharmaceutica Sinica
Year:
2013
Document type:
Article