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KMID : 0545120100200101430
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Journal of Microbiology and Biotechnology
2010 Volume.20 No. 10 p.1430 ~ p.1435
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The Orientation-dependent expression of angiostatin-endostatin hybrid proteins and their characterization for the synergistic effects of antiangiogenesis
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Paek Sun-Yeol
Kim Yong-Seok
Choi Shin-Geon
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Abstract
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Two angiostatic fusion proteins (hAE and hEA) of human angiostatin (hAS) and endostatin (hES) proteins differed in tandem connection manner were constructed and evaluated for synergistic anti-angiogenic effects. The 65 kDa secreted fusion proteins from Pichia pastoris expression were verified by mass-spec analysis and western blotting assay. Luciferase reporter gene assay using VEGF promoter revealed that angiostatin-endostatin fusion protein (hAE) and its corresponding fusion gene delivery on Human Microvascular Endothelial Cells (HMEC 1) resulted in more potent synergistic anti-angiogenic effects than endostatin-angiostatin fusion protein (hEA). These facts suggest that the orientation of fusion genes between hAS and hES might be an important factor for developing therapeutic proteins.
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KEYWORD
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angiogenesis inhibitor, angiostatin, endostatin, fusion protein, Pichia expression
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