Characterization of the primary structure of TNK-tissue plasminogen activator using LC-MS.
- Author:
Lei TAO
;
Youxue DING
;
Ying GUO
;
Chunming RAO
;
Junzhi WANG
- Publication Type:Journal Article
- From:
Acta Pharmaceutica Sinica
2013;48(6):896-900
- CountryChina
- Language:Chinese
-
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.
