Abstract
A tetrapeptide, RGDS, was inserted into proUK kringle domain G118-L119 by the construction of a mutant proUK-RGDS gene. The gene was expressed in the baculovirus expression system. Immunoaffinity chromatography was used to purify the chimera and protein with purity over 90% was achieved. The chimera was tested for its platelet membrane binding function and showed a calcium-dependent platelet binding activity. Amidolytic activity of the chimera was tested. The result indicated that specific amidolytic activity of plasmin activated chimera was 62 000 IU/mg, comparable to the previously reported 65 355 IU/mg of plasmin activated natural proUK[1]. Activation of plasminogen by the chimera after plasmin treatment followed Michieal-Menten kinetics, and the Km was 0.97 μmol/L, which was also comparable to 1.64 μmol/L of native urokinase. The chimera also showed intensive ability to inhibit platelet aggregation in vitro. These results indicate that this chimera might be useful as a bifunctional thrombolytic agent.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 264-266 |
| Number of pages | 3 |
| Journal | Science in China, Series C: Life Sciences |
| Volume | 42 |
| Issue number | 3 |
| State | Published - Jun 1 1999 |
| Externally published | Yes |
Keywords
- Anti-aggregation
- Bifunctional protein
- ProUK
- RGDS
- Thrombolysis
ASJC Scopus subject areas
- General Biochemistry, Genetics and Molecular Biology
- General Environmental Science
- General Agricultural and Biological Sciences