Identification of novel candidate phosphatidic acid-binding proteins involved in the salt-stress response of Arabidopsis thaliana roots
Identification of novel candidate phosphatidic acid-binding proteins involved in the salt-stress response of Arabidopsis thaliana roots
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DOI：
10.1042/bj20121639
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发表时间：
2013-03-15
影响因子：
4.1
通讯作者：
Testerink, Christa
中科院分区：
文献类型：
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作者：
McLoughlin, Fionn;Arisz, Steven A.;Testerink, Christa
磷脂酸（PA）是一种脂质第二信使，参与植物生命周期中发生的一系列过程。这些包括发育、代谢以及生物和非生物胁迫反应。PA水平增加响应于盐，但很少有人知道它的功能在盐胁迫的最早期反应。在本研究中，我们结合了一种方法来分离拟南芥根的外周膜蛋白与脂质亲和纯化，以确定推定的蛋白质与PA相互作用，并招募到膜响应盐胁迫。在MS鉴定的42个推定的PA结合蛋白中，一组8个新的候选PA结合蛋白在盐胁迫7分钟后在膜组分处积累。其中包括CHC（网格蛋白重链）同种型、ANTH（AP 180 N-末端同源性）结构域网格蛋白组装蛋白（一种假定的钾转运调节剂）、两种核糖体蛋白、GAPDH（甘油醛3-磷酸脱氢酶）和PI（磷脂酰肌醇）4-激酶。PA结合和盐诱导的膜募集GAPDH和CHC的细胞组分的蛋白质印迹分析证实。总之，本研究的方法是一种有效的方法来分离生物学相关的脂质结合蛋白，并提供了新的线索，在PA介导的盐胁迫反应的根的研究。
PA (phosphatidic acid) is a lipid second messenger involved in an array of processes occurring during a plant's life cycle. These include development, metabolism, and both biotic and abiotic stress responses. PA levels increase in response to salt, but little is known about its function in the earliest responses to salt stress. In the present study we have combined an approach to isolate peripheral membrane proteins of Arabidopsis thaliana roots with lipid-affinity purification, to identify putative proteins that interact with PA and are recruited to the membrane in response to salt stress. Of the 42 putative PA-binding proteins identified by MS, a set of eight new candidate PA-binding proteins accumulated at the membrane fraction after 7 min of salt stress. Among these were CHC (clathrin heavy chain) isoforms, ANTH (AP180 N-terminal homology) domain clathrin-assembly proteins, a putative regulator of potassium transport, two ribosomal proteins, GAPDH (glyceraldehyde 3-phosphate dehydrogenase) and a PI (phosphatidylinositol) 4-kinase. PA binding and salt-induced membrane recruitment of GAPDH and CHC were confirmed by Western blot analysis of the cellular fractions. In conclusion, the approach of the present study is an effective way to isolate biologically relevant lipid-binding proteins and provides new leads in the study of PA-mediated salt-stress responses in roots.