Lee Hyang Yeon, Park Seung Bum
Department of Chemistry, Seoul National University, Seoul 151-747, Korea.
Mol Biosyst. 2011 Feb;7(2):304-10. doi: 10.1039/c0mb00122h. Epub 2010 Nov 16.
Small-molecule microarrays are powerful, high-throughput tools for gathering information about direct binding events between proteins of interest and small molecules. However, nonspecific binding on modified glass slides is the major factor reducing the quality of information obtained in proteomic screening with small-molecule microarrays. To improve the signal-to-noise ratio by suppressing the background signal, we tested several surface modification methods for glass slides. Jeffamine-coated slides showed a high fluorescence signal and a significantly enhanced signal-to-noise ratio. We applied this surface modification to proteomic screening of potential tyrosinase inhibitors with a small-molecule microarray and identified 2,4,4'-trihydroxychalcone as a new small-molecule binder to tyrosinase. Its actual binding and inhibitory effects on tyrosinase were validated using an SPR binding assay and an enzyme-based inhibition assay, respectively. Thus, we successfully demonstrate the application of Jeffamine-based modification to proteomics screening with small-molecule microarrays.
小分子微阵列是用于收集有关目标蛋白质与小分子之间直接结合事件信息的强大高通量工具。然而，修饰玻片上的非特异性结合是降低小分子微阵列蛋白质组学筛选中所获信息质量的主要因素。为了通过抑制背景信号来提高信噪比，我们测试了几种玻片表面修饰方法。涂有聚醚胺的玻片显示出高荧光信号和显著提高的信噪比。我们将这种表面修饰应用于用小分子微阵列进行潜在酪氨酸酶抑制剂的蛋白质组学筛选，并鉴定出2,4,4'-三羟基查尔酮是一种新的酪氨酸酶小分子结合剂。分别使用表面等离子体共振结合测定法和基于酶的抑制测定法验证了其对酪氨酸酶的实际结合和抑制作用。因此，我们成功证明了基于聚醚胺的修饰在小分子微阵列蛋白质组学筛选中的应用。
