Brandwijk Ricardo J M G E, Nesmelova Irina, Dings Ruud P M, Mayo Kevin H, Thijssen Victor L J L, Griffioen Arjan W
Angiogenesis Laboratory, Research Institute for Growth and Development, Department of Pathology, Maastricht University and University Hospital, Maastricht, The Netherlands.
Biochem Biophys Res Commun. 2005 Aug 12;333(4):1261-8. doi: 10.1016/j.bbrc.2005.06.029.
Anginex, a designed peptide 33-mer, is a potent angiogenesis inhibitor and anti-tumor agent in vivo. Anginex functions by inhibiting endothelial cell (EC) proliferation and migration leading to detachment and apoptosis of activated EC's. To better understand tumor endothelium targeting properties of anginex and enable its use in gene therapy, we constructed an artificial gene encoding the biologically exogenous peptide and produced the protein recombinantly in Pichia pastoris. Mass spectrometry shows recombinant anginex to be a dimer and circular dichroism shows the recombinant protein folds with beta-strand structure like the synthetic peptide. Moreover, like parent anginex, the recombinant protein is active at inhibiting EC growth and migration, as well as inhibiting angiogenesis in vivo in the chorioallantoic membrane of the chick embryo. This study demonstrated that it is possible to produce a functionally active protein version of a rationally designed peptide, using an artificial gene and the recombinant protein approach.
血管抑制素（Anginex）是一种设计合成的33肽，在体内是一种有效的血管生成抑制剂和抗肿瘤药物。血管抑制素通过抑制内皮细胞（EC）增殖和迁移发挥作用，导致活化的内皮细胞脱离和凋亡。为了更好地理解血管抑制素的肿瘤内皮靶向特性并使其能够用于基因治疗，我们构建了编码这种生物外源性肽的人工基因，并在毕赤酵母中重组表达该蛋白。质谱分析表明重组血管抑制素是二聚体，圆二色性显示重组蛋白折叠成具有β-链结构，类似于合成肽。此外，与天然血管抑制素一样，重组蛋白在抑制内皮细胞生长和迁移以及抑制鸡胚尿囊膜体内血管生成方面具有活性。这项研究表明，使用人工基因和重组蛋白方法有可能生产出合理设计肽的功能活性蛋白形式。
