Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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The Orientation-Dependent Expression of Angiostatin-Endostatin Hybrid Proteins and Their Characterization for the Synergistic Effects of Antiangiogenesis

  • Paek, Sun-Yeol (Department of Bioengineering and Technology, Kangwon National University) ;
  • Kim, Yong-Seok (Department of Biochemistry and Molecular Biology, College of Medicine, Hanyang University) ;
  • Choi, Shin-Geon (Department of Bioengineering and Technology, Kangwon National University)
  • Received : 2010.04.29
  • Accepted : 2010.07.05
  • Published : 2010.10.28
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Abstract

Two angiostatic fusion proteins (hAE and hEA), differing in tandem connection manners, were constructed from human angiostatin (hAS) and endostatin (hES) proteins. These fusion proteins were then evaluated for synergistic antiangiogenic properties. The 65 kDa secreted fusion proteins, expressed in Pichia pastoris, were verified by both mass analysis and Western blotting assay. Luciferase reorter gene assay, using a VEGF promoter, revealed that the angiostatin-endostatin fusion protein (hAE), and its corresponding fusion gene delivery on human microvascular endothelial cells (HMEC-1), resulted in a more potent synergistic antiangiogenic effect than the endostatin-angiostatin fusion protein (hEA). These results suggest that the orientation of the fusion genes in hAS and hES might be an important factor in the development of therapeutic proteins.

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References

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