Department of Bioengineering and Technology, Kangwon National University, Chuncheon 200-701, Korea.
J Microbiol Biotechnol. 2010 Oct;20(10):1430-5. doi: 10.4014/jmb.1004.04040.
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.
两种血管生成抑制融合蛋白（hAE 和 hEA）由人血管抑素（hAS）和内皮抑素（hES）蛋白以串联连接方式构建，并评估其协同抗血管生成作用。毕赤酵母表达的 65 kDa 分泌融合蛋白通过质谱分析和 Western blot 分析进行验证。使用 VEGF 启动子的荧光素酶报告基因检测表明，血管生成抑制素-内皮抑素融合蛋白（hAE）及其相应的融合基因在人微血管内皮细胞（HMEC-1）上的传递导致更有效的协同抗血管生成作用，强于内皮抑素-血管生成抑制素融合蛋白（hEA）。这些事实表明，hAS 和 hES 之间融合基因的方向可能是开发治疗性蛋白的一个重要因素。
