Siemeister G, Schnurr B, Mohrs K, Schächtele C, Marmé D, Martiny-Baron G
Tumor Biology Center, Institute of Molecular Medicine, Freiburg Federal Republic of Germany.
Biochem Biophys Res Commun. 1996 May 15;222(2):249-55. doi: 10.1006/bbrc.1996.0730.
Vascular endothelial growth factor (VEGF) was identified as an endothelial cell specific mitogen that induces angiogenesis and vascular permeability in vivo. VEGF is a homodimeric protein which contains three intramolecular and two intermolecular disulfide bridges. Here, we report on an efficient procedure for recombinant production of VEGF isoforms VEGF121 and VEGF165 in Escherichia coli. The proteins were solubilized from inclusion bodies, refolded, and purified by chromatographic methods. The final protein products were almost completely in the dimeric conformation, bound to VEGF receptor FLT1 with a Kd of 30 pM, stimulated proliferation of human umbilical vein endothelial cells half-maximally at a concentration of 30 pM, and induced in vivo neovascularization and vascular permeability on the chicken chorioallantoic membrane.
血管内皮生长因子（VEGF）被鉴定为一种内皮细胞特异性促有丝分裂原，可在体内诱导血管生成和血管通透性。VEGF是一种同二聚体蛋白，含有三个分子内和两个分子间二硫键。在此，我们报告了一种在大肠杆菌中重组生产VEGF亚型VEGF121和VEGF165的高效方法。这些蛋白质从包涵体中溶解、复性，并通过色谱方法纯化。最终的蛋白质产物几乎完全处于二聚体构象，以30 pM的解离常数（Kd）与VEGF受体FLT1结合，在30 pM的浓度下对人脐静脉内皮细胞的增殖产生半最大刺激，并在鸡胚绒毛尿囊膜上诱导体内新血管形成和血管通透性。
