Terzi F, Cambridge S
Institute of Cell Biology and Anatomy, University of Heidelberg, Heidelberg, Germany.
Institute of Cell Biology and Anatomy, University of Heidelberg, Heidelberg, Germany.
Methods Enzymol. 2017;585:29-47. doi: 10.1016/bs.mie.2016.09.014. Epub 2017 Jan 12.
Comparative, quantitative mass spectrometry of proteins provides great insight to protein abundance and function, but some molecular characteristics related to protein dynamics are not so easily obtained. Because the metabolic incorporation of stable amino acid isotopes allows the extraction of distinct temporal and spatial aspects of protein dynamics, the SILAC methodology is uniquely suited to be adapted for advanced labeling strategies. New SILAC strategies have emerged that allow deeper foraging into the complexity of cellular proteomes. Here, we review a few advanced SILAC-labeling strategies that have been published during last the years. Among them, different subsaturating-labeling as well as dual-labeling schemes are most prominent for a range of analyses including those of neuronal proteomes, secretion, or cell-cell-induced stimulations. These recent developments suggest that much more information can be gained from proteomic analyses if the labeling strategies are specifically tailored toward the experimental design.
蛋白质的比较定量质谱分析为蛋白质丰度和功能提供了深入见解，但一些与蛋白质动力学相关的分子特征却不那么容易获得。由于稳定氨基酸同位素的代谢掺入能够提取蛋白质动力学不同的时间和空间方面的信息，因此稳定同位素标记氨基酸细胞培养技术（SILAC）方法特别适合用于先进的标记策略。新的SILAC策略已经出现，能够更深入地探究细胞蛋白质组的复杂性。在这里，我们回顾了过去几年中发表的一些先进的SILAC标记策略。其中，不同的亚饱和标记以及双标记方案在一系列分析中最为突出，包括对神经元蛋白质组、分泌或细胞间诱导刺激的分析。这些最新进展表明，如果标记策略是根据实验设计专门定制的，那么蛋白质组学分析可以获得更多信息。
