Tao Zhihua, Ren Haiyun
Institute of Cell Biology, College of Life Science, Beijing Normal University, 100875, Beijing, China.
Sci China C Life Sci. 2003 Aug;46(4):379-88. doi: 10.1007/BF03192581.
The effect of plasma gelsolin on plant microfilaments and its localization in plant cells were investigated. The results by using ultracentrifugation and electron microscopy showed that plant microfilaments could be severed into shorter fragments by gelsolin in a Ca(2+)-dependent manner. By measuring the binding ability of plasma gelsolin to pollen actin using the method of immunoprecipitation, it was shown that pollen actin could bind gelsolin at a ratio of 2.0±0.21 in the presence of Ca(2+). Addition of EGTA could disassociate the actin-gelsolin complexes, reducing the ratio to 1.2±0.23, and the addition of PIP(2) could further reduce the ratio to 0.8±0.1. The results indicate that plant actin has similar binding properties with plasma gelsolin as that of animal actin. By Western blotting we identified the existence of gelsolin in lily pollen. The results of immunolocalization of gelsolin in pollen and pollen tube showed that gelsolin was mainly localized at the germinal furrow in pollen grains and at the cytoplasm in pollen tube, especially in the tip region.
研究了血浆凝溶胶蛋白对植物微丝的作用及其在植物细胞中的定位。超速离心和电子显微镜观察结果表明,植物微丝可被凝溶胶蛋白以依赖Ca(2+)的方式切断成较短片段。通过免疫沉淀法测定血浆凝溶胶蛋白与花粉肌动蛋白的结合能力,结果表明在Ca(2+)存在下,花粉肌动蛋白与凝溶胶蛋白的结合比例为2.0±0.21。加入EGTA可使肌动蛋白-凝溶胶蛋白复合物解离,比例降至1.2±0.23,加入PIP(2)可进一步将比例降至0.8±0.1。结果表明,植物肌动蛋白与血浆凝溶胶蛋白的结合特性与动物肌动蛋白相似。通过蛋白质免疫印迹法,我们鉴定了百合花粉中凝溶胶蛋白的存在。凝溶胶蛋白在花粉和花粉管中的免疫定位结果表明,凝溶胶蛋白主要定位于花粉粒的萌发沟和花粉管的细胞质中,尤其是顶端区域。