AtGRIP protein locates to the secretory vesicles of trans Golgi-network in Arabidopsis root cap cells
AtGRIP protein locates to the secretory vesicles of trans Golgi-network in Arabidopsis root cap cells
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DOI：
10.1007/s11434-008-0420-4
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发表时间：
2008-10-01
影响因子：
--
通讯作者：
Li Yan
中科院分区：
文献类型：
--
作者：
Chen Ying;Zhang Wei;Li Yan
GRIP 结构域蛋白位于跨高尔基体网络，被认为在酵母和动物细胞的高尔基体运输中发挥重要作用。在本研究中，AtGRIP cDNA 通过逆转录酶 PCR 从拟南芥幼苗分离的 RNA 中扩增。亲和纯化AtGRIP的GST融合蛋白，获得兔多克隆抗体。用纯化的抗AtGRIP多克隆抗体进行免疫印迹表明AtGRIP蛋白的分子量约为92 kD，并且其在拟南芥中的表达不具有组织特异性。免疫荧光标记和共聚焦显微镜显示 AtGRIP 蛋白与拟南芥根细胞中的高尔基体堆栈共定位。免疫金标记和电镜观察表明AtGRIP蛋白主要定位于拟南芥根帽细胞反式高尔基体网络分泌囊泡膜上。综上所述，这些结果表明 GRIP 结构域蛋白在植物和动物细胞之间的定位是保守的。这些结果还表明 AtGRIP 可能参与调节植物细胞中高尔基体相关囊泡的形成或分选。
GRIP domain proteins, locating to the trans-Golgi network, are thought to play an essential role in Golgi apparatus trafficking in yeast and animal cells. In the present study, AtGRIP cDNA was amplified by reverse transcriptase PCR from RNA isolated from Arabidopsis seedling. The GST fusion protein of AtGRIP was affinity-purified and its rabbit polyclonal antibody was obtained. Immuno-blotting with the purified anti-AtGRIP polyclonal antibody demonstrated that the molecular mass of AtGRIP protein is about 92 kD, and its expression is not tissue-specific in Arabidopsis. Immunoflourescent labeling and confocal microscopy revealed that the AtGRIP protein was co-localized with Golgi stacks in Arabidopsis root cells. Immuno-gold labeling and electron microscopy observation showed that AtGRIP protein was mainly located to the membrane of the secretory vesicles of trans-Golgi network in Arabidopsis root cap cells. Taken together, these results indicate that the localization of GRIP domain proteins between plants and animal cells are conserved. These results also suggest that the AtGRIP may be involved in regulating the formation or sorting of Golgi-associated vesicles in plant cells.