Protein-ligand interaction is one of the highlights of molecular recognition. The most popular application of this type of interaction is drug development which requires a high throughput screening of a ligand that binds to the target protein. Our goal was to find a binding ligand with a simple detection, and once this type of ligand was found, other methods could then be used to measure the detailed kinetic or thermodynamic parameters. We started with the idea that the ligand NMR signal would disappear if it was bound to the non-tumbling mass. In order to create the non-tumbling mass, we tried the aggregates of a target protein, which was fused to the elastin-like polypeptide. We chose the maltose binding proteinas a test case, and we tried it with several sugars, which included maltose, glucose, sucrose, lactose, galactose, maltotriose, and β-cyclodextrin. The maltose signal in the H-1 NMR spectrum disappeared completely as hoped around the protein to ligand ratio of 1:3 at 298 K where the proteins aggregated. The protein signals also disappeared upon aggregation except for the fast-moving part, which resulted in a cleaner background than the monomeric form. Since we only needed to look for a disappearing signal amongst those from the mixture, it should be useful in high throughput screening. Other types of sugars except for the maltotriose and β-cyclodextrin, which are siblings of the maltose, did not seem to bind at all. We believe that our system would be especially more effective when dealing with a smaller target protein, so both the protein and the bound ligand would lose their signals only when the aggregates formed. We hope that our proposed method would contribute to accelerating the development of the potent drug candidates by simultaneously identifying several binders directly from a mixture.

蛋白质-配体相互作用是分子识别的亮点之一。这种类型的相互作用最流行的应用是药物开发，这需要对与靶蛋白结合的配体进行高通量筛选。我们的目标是通过简单的检测找到结合配体，一旦找到这种类型的配体，就可以使用其他方法来测量详细的动力学或热力学参数。我们从这样的想法开始，即如果配体 NMR 信号与非翻转物质结合，它就会消失。为了产生不翻滚的物质，我们尝试了一种目标蛋白的聚集体，它与弹性蛋白样多肽融合。我们选择了麦芽糖结合蛋白作为测试用例，我们尝试了几种糖，包括麦芽糖、葡萄糖、蔗糖、乳糖、半乳糖、麦芽三糖、和β-环糊精。H-1 NMR 光谱中的麦芽糖信号完全消失，正如所希望的那样，在 298 K 时蛋白质与配体的比率为 1:3，蛋白质聚集的地方。除了快速移动的部分外，蛋白质信号在聚集时也消失了，这导致比单体形式更干净的背景。因为我们只需要从混合物中寻找消失的信号，它应该在高通量筛选中很有用。除了麦芽糖的同胞麦芽三糖和β-环糊精外，其他类型的糖似乎根本不结合。我们相信我们的系统在处理较小的靶蛋白时会特别有效，因此蛋白质和结合的配体只有在聚集体形成时才会失去信号。