Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of Science, Palacký University, Šlechtitelů, 2778371 Olomouc, Czech Republic.
Institute for Genomics, Biocomputing & Biotechnology, Mississippi Agricultural and Forestry Experiment Station, Mississippi State University, Starkville, MS 39759, USA.
Int J Mol Sci. 2018 Dec 26;20(1):82. doi: 10.3390/ijms20010082.
Phospholipase Dα1 (PLDα1) belongs to phospholipases, a large phospholipid hydrolyzing protein family. PLDα1 has a substrate preference for phosphatidylcholine leading to enzymatic production of phosphatidic acid, a lipid second messenger with multiple cellular functions. PLDα1 itself is implicated in biotic and abiotic stress responses. Here, we present a shot-gun differential proteomic analysis on roots of two Arabidopsis mutants compared to the wild type. Interestingly, PLDα1 deficiency leads to altered abundances of proteins involved in diverse processes related to membrane transport including endocytosis and endoplasmic reticulum-Golgi transport. PLDα1 may be involved in the stability of attachment sites of endoplasmic reticulum to the plasma membrane as suggested by increased abundance of synaptotagmin 1, which was validated by immunoblotting and whole-mount immunolabelling analyses. Moreover, we noticed a robust abundance alterations of proteins involved in mitochondrial import and electron transport chain. Notably, the abundances of numerous proteins implicated in glucosinolate biosynthesis were also affected in mutants. Our results suggest a broader biological involvement of PLDα1 than anticipated thus far, especially in the processes such as endomembrane transport, mitochondrial protein import and protein quality control, as well as glucosinolate biosynthesis.
磷脂酶 Dα1（PLDα1）属于磷脂酶，是一种大型的磷脂水解蛋白家族。PLDα1 对磷脂酰胆碱具有底物偏好性，导致酶促产生磷脂酸，这是一种具有多种细胞功能的脂质第二信使。PLDα1 本身参与生物和非生物胁迫反应。在这里，我们对与野生型相比的两个拟南芥突变体的根进行了鸟枪法差异蛋白质组学分析。有趣的是，PLDα1 缺乏会导致与膜运输相关的多种过程中涉及的蛋白质丰度发生改变，包括内吞作用和内质网-高尔基体运输。PLDα1 可能参与内质网与质膜附着位点的稳定性，这是由突触结合蛋白 1 的丰度增加所提示的，通过免疫印迹和全 mounting 免疫标记分析进行了验证。此外，我们注意到参与线粒体输入和电子传递链的蛋白质的丰度发生了明显的改变。值得注意的是，突变体中也影响了许多参与硫代葡萄糖苷生物合成的蛋白质的丰度。我们的结果表明 PLDα1 的生物学参与范围比预期的要广泛，特别是在诸如内质网运输、线粒体蛋白输入和蛋白质质量控制以及硫代葡萄糖苷生物合成等过程中。
