Rodriguez-Furlan Cecilia, Zhang Chunhua, Raikhel Natasha, Hicks Glenn R
Center for Plant Cell Biology, Institute for Integrative Genome Biology, and Department of Botany and Plant Sciences, University of California, Riverside, California.
Department of Botany and Plant Pathology, Purdue University, West Lafayette, Indiana.
Curr Protoc Plant Biol. 2017 Dec;2(4):370-378. doi: 10.1002/cppb.20062.
Target identification remains a challenging step in plant chemical genomics approaches. Drug affinity responsive target stability (DARTS) represents a straightforward technique to identify small molecules' protein targets and assist in the characterization of interactions between small molecules and putative targets identified by other methods. When a small molecule interacts with a protein, it has the potential to stabilize the protein's structure, resulting in a reduced susceptibility to protease action. During the DARTS procedure, protein extracts are treated with proteolytic enzymes, and only proteins that bind to the small molecule are protected from proteolysis. DARTS represents a protocol independent of the molecule's mechanism of action or chemical structure. Another advantage of DARTS is that it does not require additional modifications or tagging of the small molecule. The protocols outlined in this article describe in detail the DARTS technique applied to plant proteins and propose several detection procedures according to protein abundance. © 2017 by John Wiley & Sons, Inc.
在植物化学基因组学方法中，靶点识别仍然是一个具有挑战性的步骤。药物亲和力响应靶点稳定性（DARTS）是一种直接的技术，用于识别小分子的蛋白质靶点，并有助于表征小分子与通过其他方法鉴定的假定靶点之间的相互作用。当小分子与蛋白质相互作用时，它有可能稳定蛋白质的结构，从而降低对蛋白酶作用的敏感性。在DARTS实验过程中，蛋白质提取物用蛋白水解酶处理，只有与小分子结合的蛋白质能免受蛋白水解作用。DARTS是一种独立于分子作用机制或化学结构的实验方案。DARTS的另一个优点是它不需要对小分子进行额外的修饰或标记。本文概述的实验方案详细描述了应用于植物蛋白的DARTS技术，并根据蛋白质丰度提出了几种检测方法。© 2017约翰威立国际出版公司
