Seminara Anna B, Turdiev Asan, Turdiev Husan, Lee Vincent T
Department of Cell Biology and Molecular Genetics, University of Maryland at College Park, College Park, Maryland.
Curr Protoc Mol Biol. 2019 Apr;126(1):e84. doi: 10.1002/cpmb.84. Epub 2018 Dec 3.
Protein interactions with nucleic acids are important for the synthesis, regulation, and stability of macromolecules. While a number of assays are available for interrogating these interactions, the differential radial capillary action of ligand assay (DRaCALA) has been developed as an easy and flexible platform that allows for the study of individual interactions when carrying out high-throughput screening for novel binding proteins and small molecule inhibitors. In this article, we describe the principle of DRaCALA and methods that utilize DRaCALA to determine the affinity and specificity of individual protein-nucleic acid interactions as well as uses for screening for binding proteins and chemical inhibitors. © 2018 by John Wiley & Sons, Inc.
蛋白质与核酸的相互作用对于大分子的合成、调控及稳定性至关重要。虽然有多种检测方法可用于研究这些相互作用,但配体差异径向毛细管作用分析(DRaCALA)已被开发成为一个简便且灵活的平台,在对新型结合蛋白和小分子抑制剂进行高通量筛选时,它能够用于研究单个相互作用。在本文中,我们描述了DRaCALA的原理以及利用DRaCALA来确定单个蛋白质 - 核酸相互作用的亲和力和特异性的方法,还有用于筛选结合蛋白和化学抑制剂的用途。© 2018 约翰威立国际出版公司