Institute of Chemistry, Humboldt-Universität zu Berlin, Brook-Taylor-Strasse 2, 12489, Berlin, Germany.
Institute of Biology, Humboldt-Universität zu Berlin, Invalidenstrasse 42, 10115, Berlin, Germany.
Chembiochem. 2019 Jan 18;20(2):159-165. doi: 10.1002/cbic.201800643. Epub 2019 Jan 9.
The surfaces of influenza A virus (IAV) particles are packed with hundreds of homo-trimeric hemagglutinins (HAs). Monovalent sugars have low affinity for HA, but distance-optimized bivalent sialyl-LacNAc (SLN) conjugates bind it with 10 -fold enhanced potency. Herein, we describe the oligomerization of distance-optimized bivalent binders by branched and linear hybridization on long repetitive DNA templates. The most effective complexes fully inhibited IAVs at a DNA template concentration of 10 m. Although a 10 m concentration of free trisaccharide ligand is required for full inhibition of the virus, DNA templating enables a 10 -fold reduction in the amount of sugar required. Notably, hybridization-induced rigidification of the DNA templates increased the serospecificity. Cryo-TEM analysis revealed that both spaghetti-type linear forms and cotton-ball-like clusters are able to bridge several adjacent HA molecules on the IAV surface. Programmed self-assembly of ligand-nucleic acid conjugates on long DNA templates might provide generic access to target-specific, high-affinity binders of proteins on globular objects such as cells and viruses.
流感病毒（IAV）粒子的表面排列着数百个同源三聚体血凝素（HAs）。单价糖对 HA 的亲和力较低，但经过距离优化的二价唾液酸化乳糖-N-乙酰氨基葡萄糖（SLN）缀合物的结合能力增强了 10 倍。在此，我们描述了通过在长重复 DNA 模板上分支和线性杂交使距离优化的二价结合物进行寡聚化。最有效的复合物在 DNA 模板浓度为 10 m 时完全抑制了 IAV。尽管游离三糖配体需要 10 m 的浓度才能完全抑制病毒，但 DNA 模板化使所需糖的量减少了 10 倍。值得注意的是，杂交诱导的 DNA 模板的刚性增加了血清特异性。冷冻-透射电镜分析显示，两种线状形式（意大利面条型和棉花球型）都能够在 IAV 表面桥接几个相邻的 HA 分子。配体-核酸缀合物在长 DNA 模板上的程序化自组装可能为针对细胞和病毒等球形物体上的蛋白质的靶向特异性、高亲和力结合物提供通用方法。
