Label-free Quantitative Proteomics Analysis of Etiolated Maize Seedling Leaves during Greening
Label-free Quantitative Proteomics Analysis of Etiolated Maize Seedling Leaves during Greening
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DOI：
10.1074/mcp.m900187-mcp200
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发表时间：
2009-11-01
影响因子：
7
通讯作者：
Wang, Bai-Chen
中科院分区：
文献类型：
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作者：
Shen, Zhuo;Li, Ping;Wang, Bai-Chen
为了更好地了解 C-4 植物玉米发育的光调节，我们使用基于纳米级超高效液相色谱-ESI-MSE 的无标记定量蛋白质组学方法研究了绿色幼苗（对照）、黄化幼苗和光照 6 或 12 小时的黄化幼苗之间的动态蛋白质组差异。在已鉴定的 400 多种蛋白质中，有 73 种在黄化玉米幼苗绿化过程中发生了显着改变。在这 73 种蛋白质中，有 25 种被鉴定为膜蛋白，而这些膜蛋白很少用二维电泳方法鉴定，这表明我们的无标记方法在膜蛋白鉴定方面的强大功能； 31个与叶绿素生物合成、光合作用和光合碳同化的光反应有关。光系统II亚基的表达对光高度敏感；其中大多数在黄化玉米幼苗中未发现，但在光照后急剧增加，表明光合器官生物发生的复杂过程与从黑暗生长到光生长形态的转变相关。然而，转录分析表明，编码这些蛋白质的大多数转录本不受光调节。相比之下，参与碳同化的酶的 mRNA 和蛋白质水平受到光的严格调控。此外，磷酸烯醇丙酮酸羧激酶（磷酸烯醇丙酮酸羧激酶 C-4 途径的关键酶）比 NADP-苹果酸酶 C-4 途径的关键酶更受光调节。此外，本研究还鉴定了最初报道在根中特异性表达的磷酸烯醇丙酮酸羧化酶 1C；该酶的表达比其异构体对光更敏感。综上所述，这些结果代表了C 4 植物黄化幼苗绿化的全面动态蛋白质谱和光调控网络，为进一步研究C 4 植物光诱导发育中的基因功能和调控机制提供了基础。分子与细胞蛋白质组学 8：2443-2460，2009。
To better understand light regulation of C-4 plant maize development, we investigated dynamic proteomic differences between green seedlings (control), etiolated seedlings, and etiolated seedlings illuminated for 6 or 12 h using a label-free quantitative proteomics approach based on nanoscale ultraperformance liquid chromatography-ESI-MSE. Among more than 400 proteins identified, 73 were significantly altered during etiolated maize seedling greening. Of these 73 proteins, 25 were identified as membrane proteins that seldom had been identified with two-dimensional electrophoresis methods, indicating the power of our label-free method for membrane protein identification; 31 were related to light reactions of chlorophyll biosynthesis, photosynthesis, and photosynthetic carbon assimilation. The expression of photosystem II subunits was highly sensitive to light; most of them were not identified in etiolated maize seedlings but drastically increased upon light exposure, indicating that the complex process of biogenesis of the photosynthetic apparatus correlates with the transition from a dark-grown to a light-grown morphology. However, transcriptional analysis indicated that most transcripts encoding these proteins were not regulated by light. In contrast, the levels of mRNAs and proteins for enzymes involved in carbon assimilation were tightly regulated by light. Additionally phosphoenolpyruvate carboxykinase, the key enzyme of the phosphoenolpyruvate carboxykinase C-4 pathway, was more tightly regulated by light than the key enzymes of the NADP-malic enzyme C-4 pathway. Furthermore phosphoenolpyruvate carboxylase 1C, which was originally reported to be specifically expressed in roots, was also identified in this study; expression of this enzyme was more sensitive to light than its isoforms. Taken together, these results represent a comprehensive dynamic protein profile and light-regulated network of C 4 plants for etiolated seedling greening and provide a basis for further study of the mechanism of gene function and regulation in light-induced development of C-4 plants. Molecular & Cellular Proteomics 8: 2443-2460, 2009.