Drug Affinity Responsive Target Stability (DARTS) to Resolve Protein–Small Molecule Interaction in Arabidopsis

Q1 Agricultural and Biological Sciences

Current protocols in plant biology Pub Date : 2018-02-13 DOI:10.1002/cppb.20062

Cecilia Rodriguez-Furlan, Chunhua Zhang, Natasha Raikhel, Glenn R. Hicks

{"title":"Drug Affinity Responsive Target Stability (DARTS) to Resolve Protein–Small Molecule Interaction in Arabidopsis","authors":"Cecilia Rodriguez-Furlan, Chunhua Zhang, Natasha Raikhel, Glenn R. Hicks","doi":"10.1002/cppb.20062","DOIUrl":null,"url":null,"abstract":"<p>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.</p>","PeriodicalId":10932,"journal":{"name":"Current protocols in plant biology","volume":"2 4","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2018-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cppb.20062","citationCount":"5","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current protocols in plant biology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cppb.20062","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Agricultural and Biological Sciences","Score":null,"Total":0}

引用次数: 5

Abstract

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.

查看原文本刊更多论文

拟南芥蛋白-小分子相互作用的药物亲和反应靶稳定性(DARTS

在植物化学基因组学方法中，目标鉴定仍然是一个具有挑战性的步骤。药物亲和反应性靶标稳定性(DARTS)是一种简单的技术，可以识别小分子的蛋白质靶标，并有助于表征小分子与其他方法确定的假定靶标之间的相互作用。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易感性。在DARTS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才不会被蛋白质水解。DARTS代表了一种独立于分子作用机制或化学结构的协议。dart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。本文概述的方案详细描述了DARTS技术在植物蛋白中的应用，并根据蛋白质丰度提出了几种检测方法。©2017 by John Wiley &儿子,Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Current protocols in plant biology Agricultural and Biological Sciences-Plant Science

自引率

0.00%

发文量

期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. Current Protocols in Plant Biology provides reproducible step-by-step instructions for protocols relevant to plant research. Furthermore, Current Protocols content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols in Plant Biology to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Plant Biology is the comprehensive source for protocols in the multidisciplinary field of plant biology, providing an extensive range of protocols from basic to cutting edge. Coverage includes: Extraction and analysis of DNA, RNA, proteins Chromosome analysis Transcriptional analysis Protein expression Metabolites Plant enzymology Epigenetics Plant genetic transformation Mutagenesis Arabidopsis, Maize, Poplar, Rice, and Soybean, and more.