Laboratorio NMR, Istituto per lo Studio delle Macromolecole, Consiglio Nazionale delle Ricerche, Milano, Italy.
PLoS One. 2012;7(5):e36990. doi: 10.1371/journal.pone.0036990. Epub 2012 May 14.
Fibroblast growth factors (FGFs) are recognized targets for the development of therapies against angiogenesis-driven diseases, including cancer. The formation of a ternary complex with the transmembrane tyrosine kinase receptors (FGFRs), and heparan sulphate proteoglycans (HSPGs) is required for FGF2 pro-angiogenic activity. Here by using a combination of techniques including Nuclear Magnetic Resonance, Molecular Dynamics, Surface Plasmon Resonance and cell-based binding assays we clarify the molecular mechanism of inhibition of an angiostatic small molecule, sm27, mimicking the endogenous inhibitor of angiogenesis, thrombospondin-1. NMR and MD data demonstrate that sm27 engages the heparin-binding site of FGF2 and induces long-range dynamics perturbations along FGF2/FGFR1 interface regions. The functional consequence of the inhibitor binding is an impaired FGF2 interaction with both its receptors, as demonstrated by SPR and cell-based binding assays. We propose that sm27 antiangiogenic activity is based on a twofold-direct and allosteric-mechanism, inhibiting FGF2 binding to both its receptors.
成纤维细胞生长因子（FGFs）是针对血管生成驱动的疾病（包括癌症）开发治疗方法的公认靶点。与跨膜酪氨酸激酶受体（FGFRs）和硫酸乙酰肝素蛋白聚糖（HSPGs）形成三元复合物是 FGF2 促血管生成活性所必需的。在这里，我们通过结合使用包括核磁共振、分子动力学、表面等离子体共振和基于细胞的结合测定在内的一系列技术，阐明了模仿内源性血管生成抑制剂——血栓素-1 的血管生成抑制小分子 sm27 的抑制机制。NMR 和 MD 数据表明，sm27 与 FGF2 的肝素结合位点结合，并诱导 FGF2/FGFR1 界面区域的长程动力学扰动。SPR 和基于细胞的结合测定表明，抑制剂结合的功能后果是 FGF2 与其两种受体的相互作用受损。我们提出，sm27 的抗血管生成活性基于双重直接和变构机制，抑制 FGF2 与其两种受体的结合。
