Plant SILAC: stable-isotope labelling with amino acids of arabidopsis seedlings for quantitative proteomics.
Plant SILAC: stable-isotope labelling with amino acids of arabidopsis seedlings for quantitative proteomics.
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DOI：
10.1371/journal.pone.0072207
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Brown JW
中科院分区：
文献类型：
--
作者：
Lewandowska D;ten Have S;Hodge K;Tillemans V;Lamond AI;Brown JW
细胞培养中氨基酸稳定同位素标记 (SILAC) 是一种用于比较定量蛋白质组学的强大技术，最近已应用于许多不同的真核生物。拟南芥细胞培养物中标记氨基酸的低效掺入导致 SILAC 在植物系统中的使用非常有限。我们提出了一种方法，首次可以对整个拟南芥幼苗的含稳定同位素的精氨酸和赖氨酸进行有效标记。为了说明该方法的实用性，我们将高标记效率（> 95％）与暴露于增加的盐浓度的幼苗的定量蛋白质组学分析结合起来。在用 80 mM NaCl（相对温和的盐胁迫）处理 7 天的植物中，鉴定出 215 个蛋白质，其表达水平与未处理的幼苗对照相比发生显着变化。 92 个上调的蛋白质包括参与非生物应激反应和光合作用的蛋白质，而 123 个下调的蛋白质分别富含参与减少氧化应激和其他应激反应的蛋白质。通过这种改进的 SILAC 方法对整个拟南芥幼苗进行有效标记，为开发拟南芥遗传资源和分析突变对体内定量蛋白质动态的影响提供了新的机会。
Stable Isotope Labelling by Amino acids in Cell culture (SILAC) is a powerful technique for comparative quantitative proteomics, which has recently been applied to a number of different eukaryotic organisms. Inefficient incorporation of labelled amino acids in cell cultures of Arabidopsis thaliana has led to very limited use of SILAC in plant systems. We present a method allowing, for the first time, efficient labelling with stable isotope-containing arginine and lysine of whole Arabidopsis seedlings. To illustrate the utility of this method, we have combined the high labelling efficiency (>95%) with quantitative proteomics analyses of seedlings exposed to increased salt concentration. In plants treated for 7 days with 80 mM NaCl, a relatively mild salt stress, 215 proteins were identified whose expression levels changed significantly compared to untreated seedling controls. The 92 up-regulated proteins included proteins involved in abiotic stress responses and photosynthesis, while the 123 down-regulated proteins were enriched in proteins involved in reduction of oxidative stress and other stress responses, respectively. Efficient labelling of whole Arabidopsis seedlings by this modified SILAC method opens new opportunities to exploit the genetic resources of Arabidopsis and analyse the impact of mutations on quantitative protein dynamics in vivo.
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影响因子：
3.4
作者：
Naumann, Bianca;Busch, Andreas;Hippler, Michael
通讯作者：
Hippler, Michael
影响因子：
4.4
作者：
Cox, Juergen;Neuhauser, Nadin;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
3.4
作者：
Lanquar, Viviane;Kuhn, Lauriane;Thomine, Sebastien
通讯作者：
Thomine, Sebastien
影响因子：
4.4
作者：
Mastrobuoni G;Irgang S;Pietzke M;Assmus HE;Wenzel M;Schulze WX;Kempa S
通讯作者：
Kempa S
用K（15）NO3作为蛋白质和代谢产物定量分析的工具，对植物细胞培养的代谢标记。
影响因子：
5.1
作者：
Engelsberger WR;Erban A;Kopka J;Schulze WX
通讯作者：
Schulze WX