Wang Wayne U, Chen Chuo, Lin Keng-hui, Fang Ying, Lieber Charles M
Department of Chemistry and Chemical Biology, Biophysics Program, and Division of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138, USA.
Proc Natl Acad Sci U S A. 2005 Mar 1;102(9):3208-12. doi: 10.1073/pnas.0406368102. Epub 2005 Feb 16.
Development of miniaturized devices that enable rapid and direct analysis of the specific binding of small molecules to proteins could be of substantial importance to the discovery of and screening for new drug molecules. Here, we report highly sensitive and label-free direct electrical detection of small-molecule inhibitors of ATP binding to Abl by using silicon nanowire field-effect transistor devices. Abl, which is a protein tyrosine kinase whose constitutive activity is responsible for chronic myelogenous leukemia, was covalently linked to the surfaces of silicon nanowires within microfluidic channels to create active electrical devices. Concentration-dependent binding of ATP and concentration-dependent inhibition of ATP binding by the competitive small-molecule antagonist STI-571 (Gleevec) were assessed by monitoring the nanowire conductance. In addition, concentration-dependent inhibition of ATP binding was examined for four additional small molecules, including reported and previously unreported inhibitors. These studies demonstrate that the silicon nanowire devices can readily and rapidly distinguish the affinities of distinct small-molecule inhibitors and, thus, could serve as a technology platform for drug discovery.
开发能够快速直接分析小分子与蛋白质特异性结合的小型化设备，对于新药分子的发现和筛选可能具有重大意义。在此，我们报告了使用硅纳米线场效应晶体管设备对ATP与Abl结合的小分子抑制剂进行高灵敏度、无标记的直接电学检测。Abl是一种蛋白质酪氨酸激酶，其组成性活性与慢性粒细胞白血病有关，它被共价连接到微流控通道内硅纳米线的表面，以制造有源电子设备。通过监测纳米线电导来评估ATP的浓度依赖性结合以及竞争性小分子拮抗剂STI-571（格列卫）对ATP结合的浓度依赖性抑制。此外，还对另外四种小分子进行了ATP结合的浓度依赖性抑制研究，包括已报道和先前未报道的抑制剂。这些研究表明，硅纳米线设备能够轻松快速地区分不同小分子抑制剂的亲和力，因此可作为药物发现的技术平台。
