A prourokinase-RGDS chimera - Construction, expression and characterization

Bin Qian, Yingqing Sun, Yan Guo, Xin Dang, Binggen Ru*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

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 languageEnglish (US)
Pages (from-to)264-266
Number of pages3
JournalScience in China, Series C: Life Sciences
Volume42
Issue number3
StatePublished - Jun 1 1999
Externally publishedYes

Keywords

  • Anti-aggregation
  • Bifunctional protein
  • ProUK
  • RGDS
  • Thrombolysis

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Environmental Science(all)
  • Agricultural and Biological Sciences(all)

Fingerprint

Dive into the research topics of 'A prourokinase-RGDS chimera - Construction, expression and characterization'. Together they form a unique fingerprint.

Cite this