用于相对蛋白质定量的NeuCode标签。
NeuCode labels for relative protein quantification.
作者信息
Merrill Anna E, Hebert Alexander S, MacGilvray Matthew E, Rose Christopher M, Bailey Derek J, Bradley Joel C, Wood William W, El Masri Marwan, Westphall Michael S, Gasch Audrey P, Coon Joshua J
机构信息
From the ‡Genome Center of Wisconsin, University of Wisconsin, Madison, Wisconsin 53706; §Department of Chemistry, University of Wisconsin, Madison, Wisconsin 53706;
From the ‡Genome Center of Wisconsin, University of Wisconsin, Madison, Wisconsin 53706;
出版信息
Mol Cell Proteomics. 2014 Sep;13(9):2503-12. doi: 10.1074/mcp.M114.040287. Epub 2014 Jun 17.
We describe a synthesis strategy for the preparation of lysine isotopologues that differ in mass by as little as 6 mDa. We demonstrate that incorporation of these molecules into the proteomes of actively growing cells does not affect cellular proliferation, and we discuss how to use the embedded mass signatures (neutron encoding (NeuCode)) for multiplexed proteome quantification by means of high-resolution mass spectrometry. NeuCode SILAC amalgamates the quantitative accuracy of SILAC with the multiplexing of isobaric tags and, in doing so, offers up new opportunities for biological investigation. We applied NeuCode SILAC to examine the relationship between transcript and protein levels in yeast cells responding to environmental stress. Finally, we monitored the time-resolved responses of five signaling mutants in a single 18-plex experiment.
我们描述了一种用于制备质量差异低至6 mDa的赖氨酸同位素异构体的合成策略。我们证明，将这些分子掺入活跃生长细胞的蛋白质组中不会影响细胞增殖，并且我们讨论了如何通过高分辨率质谱法利用嵌入的质量特征（中子编码（NeuCode））进行多重蛋白质组定量。NeuCode SILAC将SILAC的定量准确性与等压标签的多重化相结合，从而为生物学研究提供了新的机会。我们应用NeuCode SILAC来研究酵母细胞在应对环境压力时转录水平与蛋白质水平之间的关系。最后，我们在一个单一的18重实验中监测了五个信号突变体的时间分辨反应。
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