Botanisches Institut Universität Bonn, Venusbergweg 22, D-5300 Bonn 1, Federal Republic of Germany.
Plant Physiol. 1992 Aug;99(4):1635-41. doi: 10.1104/pp.99.4.1635.
The ether phospholipid platelet-activating factor and certain similar phospholipids, including lysophosphatidylcholine, are known to stimulate both H(+) transport and protein phosphorylation in plant microsomal membranes. In the present work, several polypeptides in highly purified tonoplast membranes from zucchini (Cucurbita pepo L.) showed platelet-activating factor-dependent phosphorylation. Comparison of protein phosphorylation in different membrane fractions separated by sucrose step density gradient centrifugation indicated that some of the phosphoproteins were contaminants or were common to several membrane fractions, but platelet-activating factor-dependent phosphorylation of peptides at 30, 53, and perhaps 100 kilodaltons was tonoplast specific. The phosphoprotein of 53 kilodaltons was shown by three different approaches (one- and two-dimensional polyacrylamide gel electrophoresis, western blots, and immunoprecipitation) to cross-react with antibody raised against the B subunit of the tonoplast ATPase from red beet (Beta vulgaris L.).
醚磷脂血小板激活因子和某些类似的磷脂，包括溶血磷脂酰胆碱，已知可刺激植物微粒体膜中的 H(+)转运和蛋白质磷酸化。在本工作中，从南瓜（Cucurbita pepo L.）中高度纯化的液泡膜中的几种多肽表现出血小板激活因子依赖性磷酸化。通过蔗糖分步密度梯度离心分离的不同膜部分的蛋白质磷酸化比较表明，一些磷酸蛋白是污染物或存在于几种膜部分中，但 30、53 和可能 100 千道尔顿的肽的血小板激活因子依赖性磷酸化是液泡膜特异性的。通过三种不同的方法（一维和二维聚丙烯酰胺凝胶电泳、western 印迹和免疫沉淀）证明 53 千道尔顿的磷酸蛋白与从红甜菜（Beta vulgaris L.）中提取的液泡膜 ATP 酶 B 亚基抗体发生交叉反应。
