蛋白质水解
计算生物学
化学
小分子
蛋白质-蛋白质相互作用
拟南芥
蛋白酶
细胞生物学
药物发现
拟南芥
双分子荧光互补
生物化学
生物
生物物理学
配体(生物化学)
突变体
结合位点
血浆蛋白结合
酶
基因
作者
Cecilia Rodriguez-Furlan,Chunhua Zhang,Natasha V. Raikhel,Glenn R. Hicks
摘要
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.
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