Glycosystems Laboratory, Instituto de Investigaciones Químicas (IIQ), cicCartuja, CSIC and Universidad de Sevilla, Americo Vespucio, 49, 41092 Sevilla, Spain.
Molecules. 2019 Apr 22;24(8):1591. doi: 10.3390/molecules24081591.
Here, we report the synthesis of a sulfated, fully protected hexasaccharide as a glycosaminoglycan mimetic and the study of its interactions with different growth factors: midkine, basic fibroblast growth factor (FGF-2) and nerve growth factor (NGF). Following a fluorous-assisted approach, monosaccharide building blocks were successfully assembled and the target oligosaccharide was prepared in excellent yield. The use of more acid stable 4,6--silylidene protected glucosamine units was crucial for the efficiency of this strategy because harsh reaction conditions were needed in the glycosylations to avoid the formation of orthoester side products. Fluorescence polarization experiments demonstrated the strong interactions between the synthesized hexamer, and midkine and FGF-2. In addition, we have developed an alternative assay to analyse these molecular recognition events. The prepared oligosaccharide was non-covalently attached to a fluorous-functionalized microplate and the direct binding of the protein to the sugar-immobilized surface was measured, affording the corresponding K value.
在这里，我们报告了一种磺酸化的、完全保护的六糖的合成，作为糖胺聚糖模拟物，并研究了它与不同生长因子的相互作用：中期因子、碱性成纤维细胞生长因子（FGF-2）和神经生长因子（NGF）。采用氟辅助方法，成功地组装了单糖砌块，并以优异的产率制备了目标寡糖。使用更耐酸的 4,6--硅亚基保护的葡萄糖胺单元对于该策略的效率至关重要，因为在糖苷化反应中需要苛刻的反应条件，以避免形成邻酯副产物。荧光偏振实验证明了合成的六聚体与中期因子和 FGF-2 之间的强烈相互作用。此外，我们还开发了一种替代分析这些分子识别事件的方法。将制备的寡糖非共价连接到氟功能化的微孔板上，并测量蛋白质与糖固定化表面的直接结合，从而获得相应的 K 值。
