A bioassay-guided fractionation system to identify endogenous small molecules that activate plasma membrane H+-ATPase activity in Arabidopsis.
A bioassay-guided fractionation system to identify endogenous small molecules that activate plasma membrane H+-ATPase activity in Arabidopsis.
复制标题
生物测定引导的分级分离系统，用于鉴定激活拟南芥质膜 H -ATP 酶活性的内源小分子
DOI：
10.1093/jxb/erx156
复制
发表时间：
2017-05-17
影响因子：
6.9
通讯作者：
Guo Y
中科院分区：
文献类型：
--
作者：
Han X;Yang Y;Wu Y;Liu X;Lei X;Guo Y
Endogenous unsaturated fatty acids identified from salt-treated Arabidopsis by a bioassay-guided fractionation method activate plasma membrane H+-ATPase by binding to its C-terminus. Plasma membrane (PM) H+-ATPase is essential for plant growth and development. Various environmental stimuli regulate its activity, a process that involves many protein cofactors. However, whether endogenous small molecules play a role in this regulation remains unknown. Here, we describe a bio-guided isolation method to identify endogenous small molecules that regulate PM H+-ATPase activity. We obtained crude extracts from Arabidopsis seedlings with or without salt treatment and then purified them into fractions based on polarity and molecular mass by repeated column chromatography. By evaluating the effect of each fraction on PM H+-ATPase activity, we found that fractions containing the endogenous, free unsaturated fatty acids oleic acid (C18:1), linoleic acid (C18:2), and linolenic acid (C18:3) extracted from salt-treated seedlings stimulate PM H+-ATPase activity. These results were further confirmed by the addition of exogenous C18:1, C18:2, or C18:3 in the activity assay. The ssi2 mutant, with reduced levels of C18:1, C18:2, and C18:3, displayed reduced PM H+-ATPase activity. Furthermore, C18:1, C18:2, and C18:3 directly bound to the C-terminus of the PM H+-ATPase AHA2. Collectively, our results demonstrate that the binding of free unsaturated fatty acids to the C-terminus of PM H+-ATPase is required for its activation under salt stress. The bio-guided isolation model described in this study could enable the identification of new endogenous small molecules that modulate essential protein functions, as well as signal transduction, in plants.
登录
查看更多内容
DOI：
10.1126/science.1244454
发表时间：
2014-01-24
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Haruta M;Sabat G;Stecker K;Minkoff BB;Sussman MR
通讯作者：
Sussman MR
影响因子：
4.5
作者：
Duby, Geoffrey;Boutry, Marc
通讯作者：
Boutry, Marc
影响因子：
9.5
作者：
Haruta M;Gray WM;Sussman MR
通讯作者：
Sussman MR
影响因子：
11.4
作者：
Kinoshita, T;Shimazaki, K
通讯作者：
Shimazaki, K
影响因子：
11.6
作者：
Kim, Hae Jin;Ok, Sung Han;Shin, Jeong Sheop
通讯作者：
Shin, Jeong Sheop