Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina 27599, USA; email:
Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina 27599, USA.
Annu Rev Anal Chem (Palo Alto Calif). 2018 Jun 12;11(1):49-77. doi: 10.1146/annurev-anchem-061516-045357.
Mass spectrometry-based quantitative proteomics is a powerful tool for gaining insights into function and dynamics of biological systems. However, peptides with different sequences have different ionization efficiencies, and their intensities in a mass spectrum are not correlated with their abundances. Therefore, various label-free or stable isotope label-based quantitation methods have emerged to assist mass spectrometry to perform comparative proteomic experiments, thus enabling nonbiased identification of thousands of proteins differentially expressed in healthy versus diseased cells. Here, we discuss the most widely used label-free and metabolic-, enzymatic-, and chemical labeling-based proteomic strategies for relative and absolute quantitation. We summarize the specific strengths and weaknesses of each technique in terms of quantification accuracy, proteome coverage, multiplexing capability, and robustness. Applications of each strategy for solving specific biological complexities are also presented.
基于质谱的定量蛋白质组学是深入了解生物系统功能和动态的有力工具。然而，具有不同序列的肽具有不同的离子化效率，并且它们在质谱中的强度与丰度不成正比。因此，出现了各种无标记或稳定同位素标记定量方法来辅助质谱进行比较蛋白质组学实验，从而能够非偏地鉴定健康细胞与患病细胞中差异表达的数千种蛋白质。在这里，我们讨论了最广泛使用的无标记和基于代谢、酶和化学标记的蛋白质组学相对和绝对定量策略。我们根据定量准确性、蛋白质组覆盖范围、多重检测能力和稳健性总结了每种技术的具体优缺点。还介绍了每种策略在解决特定生物学复杂性方面的应用。
