Cao Y, Chen H, Zhou L, Chiang M K, Anand-Apte B, Weatherbee J A, Wang Y, Fang F, Flanagan J G, Tsang M L
Department of Surgery, Harvard Medical School, Boston, Massachusetts 02115, USA.
J Biol Chem. 1996 Feb 9;271(6):3154-62. doi: 10.1074/jbc.271.6.3154.
Here we show that the Escherichia coli expressed monomers of placenta growth factor (PLGF)129 and vascular endothelial growth factor (VEGF)165 can be re-folded in vitro to form PLGF/VEGF heterodimers. The purified recombinant PLGF/VEGF heterodimers and VEGF homodimers have potent mitogenic and chemotactic effects on endothelial cells. However, PLGF/VEGF heterodimers display 20-50-fold less mitogenic activity than VEGF165 homodimers. In contrast, PLGF129 homodimers have little or no effect in these in vitro assays. We also demonstrate the presence of natural PLGF/VEGF heterodimers in the conditioned media of various human tumor cell lines. While PLGF/VEGF heterodimers bind with high affinity to a soluble Flk-1/KDR receptor, PLGF129 homodimers fail to bind to this receptor. Cross-linking of 125I-ligands to human umbilical vein endothelial cells reveals that PLGF/VEGF heterodimers and VEGF165 homodimers, but not PLGF129 homodimers, form complexes with membrane receptors. VEGF165 homodimers and PLGF/VEGF heterodimers stimulate tyrosine phosphorylation of a 220-kDa protein, the expected size for the KDR receptor in human umbilical vein endothelial cells, whereas PLGF129 homodimers are unable to induce tyrosine phosphorylation of this protein. These data indicate that PLGF may modulate VEGF-induced angiogenesis by the formation of PLGF/VEGF heterodimers in cells producing both factors.
我们在此表明,大肠杆菌表达的胎盘生长因子(PLGF)129和血管内皮生长因子(VEGF)165单体可在体外重新折叠形成PLGF/VEGF异二聚体。纯化的重组PLGF/VEGF异二聚体和VEGF同二聚体对内皮细胞具有强大的促有丝分裂和趋化作用。然而,PLGF/VEGF异二聚体的促有丝分裂活性比VEGF165同二聚体低20至50倍。相比之下,PLGF129同二聚体在这些体外试验中几乎没有或没有作用。我们还证明了在各种人类肿瘤细胞系的条件培养基中存在天然的PLGF/VEGF异二聚体。虽然PLGF/VEGF异二聚体与可溶性Flk-1/KDR受体具有高亲和力结合,但PLGF129同二聚体无法与该受体结合。将125I配体与人脐静脉内皮细胞交联显示,PLGF/VEGF异二聚体和VEGF165同二聚体而非PLGF129同二聚体与膜受体形成复合物。VEGF165同二聚体和PLGF/VEGF异二聚体刺激一种220 kDa蛋白的酪氨酸磷酸化,这是人脐静脉内皮细胞中KDR受体的预期大小,而PLGF129同二聚体无法诱导该蛋白的酪氨酸磷酸化。这些数据表明,PLGF可能通过在同时产生这两种因子的细胞中形成PLGF/VEGF异二聚体来调节VEGF诱导的血管生成。