Molecular Characterization of Mucus Binding
Molecular Characterization of Mucus Binding
复制标题
粘液结合的分子表征
DOI：
10.1021/acs.biomac.8b01467
复制
发表时间：
2019
影响因子：
6.2
通讯作者：
Ribbeck, Katharina
中科院分区：
文献类型：
--
作者：
Witten, Jacob;Samad, Tahoura;Ribbeck, Katharina
小分子与体内粘膜的结合在人类健康中具有重要作用，因为它可以影响在粘膜环境中起作用的任何分子的扩散性和活性。药物和毒素与来自粘膜病原体的信号分子的结合具有特别的临床意义。尽管粘液-小分子结合的重要性，但缺乏揭示导致粘液结合的小分子的精确化学特征的数据。我们开发了一种新的平衡透析测定法来测量小分子文库与粘蛋白和其他粘液组分的结合，从而大幅提高了小分子结合测量的通量。我们通过定量抗生素黏菌素与粘蛋白的结合来验证我们方法的生物相关性，并显示这种结合与黏菌素生物活性的抑制相关。接下来，我们使用小分子微阵列来鉴定2，4_二氨基嘧啶作为粘蛋白结合基序，并使用平衡透析确认该基序对于粘蛋白结合的重要性。此外，我们发现，对于具有这种基序的分子，与粘蛋白和粘液相关的生物聚合物DNA和藻酸盐的结合受到疏水性和电荷差异的调节。最后，我们发现，缺乏基序的分子表现出不同的结合趋势，从那些含有基序。这些结果开辟了小分子与粘液结合的常规测试和针对临床相关粘液结合性质的药物优化的前景。
Binding of small molecules to mucus membranes in the body has an important role in human health, as it can affect the diffusivity and activity of any molecule that acts in a mucosal environment. The binding of drugs and of toxins and signaling molecules from mucosal pathogens is of particular clinical interest. Despite the importance of mucus–small molecule binding, there is a lack of data revealing the precise chemical features of small molecules that lead to mucus binding. We developed a novel equilibrium dialysis assay to measure the binding of libraries of small molecules to mucin and other mucus components, substantially increasing the throughput of small molecule binding measurements. We validated the biological relevance of our approach by quantifying binding of the antibiotic colistin to mucin, and showing that this binding was associated with inhibition of colistin’s bioactivity. We next used a small molecule microarray to identify 2,4-diaminopyrimidine as a mucin binding motif and confirmed the importance of this motif for mucin binding using equilibrium dialysis. Furthermore, we showed that, for molecules with this motif, binding to mucins and the mucus-associated biopolymers DNA and alginate is modulated by differences in hydrophobicity and charge. Finally, we showed that molecules lacking the motif exhibited different binding trends from those containing the motif. These results open up the prospect of routine testing of small molecule binding to mucus and optimization of drugs for clinically relevant mucus binding properties.