Proteomic Analysis of Rice Endosperm Cells in Response to Expression of hGM-CSF
Proteomic Analysis of Rice Endosperm Cells in Response to Expression of hGM-CSF
复制标题
水稻胚乳细胞对 hGM-CSF 表达的响应的蛋白质组学分析
DOI：
10.1021/pr8002968
复制
发表时间：
2009-02-01
影响因子：
4.4
通讯作者：
Yang, Daichang
中科院分区：
文献类型：
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作者：
Luo, Junling;Ning, Tingting;Yang, Daichang
转基因产物在植物细胞中的大量积累不仅是作物改良所必需的，而且也是分子嫁接所必需的。然而，对转基因产物和内源蛋白质在谷物细胞中的命运的知识是缺乏的。在这里，我们利用定量质谱为基础的蛋白质组学方法的表达谱的转基因水稻胚乳细胞的表达响应于表达的重组药物蛋白，人粒细胞-巨噬细胞集落刺激因子（hGM-CSF）的比较分析。本研究首次提供了有关外源和内源蛋白质在籽粒细胞中命运的证据。在假阳性率为5%的1883个鉴定的蛋白质中，103个在转基因和野生型胚乳细胞之间显示出显著变化（p值< 0.05）。在转基因水稻胚乳中，内源贮藏蛋白和大多数碳水化合物代谢相关蛋白表达下调，而26 S蛋白酶体相关蛋白和分子伴侣表达上调。此外，我们观察到hGM-CSF的表达诱导内质网应激并激活泛素/26 S-蛋白酶体途径，这导致该外源基因产物在转基因水稻胚乳中的泛素化。
The accumulation of significant levels of transgenic products in plant cells is required not only for crop improvement, but also for molecular pharming. However, knowledge about the fate of transgenic products and endogenous proteins in grain cells is lacking. Here, we utilized a quantitative mass spectrometry-based proteomic approach for comparative analysis of expression profiles of transgenic rice endosperm cells in response to expression of a recombinant pharmaceutical protein, human granulocyte-macrophage colony stimulation factor (hGM-CSF). This study provided the first available evidence concerning the fate of exogenous and endogenous proteins in grain cells. Among 1883 identified proteins with a false positive rate of 5%, 103 displayed significant changes (p-value < 0.05) between the transgenic and the wild-type endosperm cells. Notably, endogenous storage proteins and most carbohydrate metabolism-related proteins were down-regulated, while 26S proteasome-related proteins and chaperones were up-regulated in the transgenic rice endosperm. Furthermore, it was observed that expression of hGM-CSF induced endoplasmic reticulum stress and activated the ubiquitin/26S-proteasome pathway, which led to ubiquitination of this foreign gene product in the transgenic rice endosperm.