Emadali Anouk, Gallagher-Gambarelli Maighread
CEA, DSV, iRTSV, Laboratoire d'étude de dynamique des protéomes, Grenoble, F-38054, France.
Med Sci (Paris). 2009 Oct;25(10):835-42. doi: 10.1051/medsci/20092510835.
Mass spectrometry-based quantitative proteomics strategies are ideally adapted to the detection of global protein changes between different biological samples. Among these, SILAC (stable isotope labelling by amino acids in cell culture) has demonstrated a great potential. This method is extremely accurate and relatively easy to apply for the quantification of proteins extracted from cultured cells. SILAC involves cell culture either in regular culture media ("light" protein synthesis) or in media where amino acids have been replaced by their isotopically labelled counterparts ("heavy" protein synthesis). Cell populations to be compared can be mixed and treated as a single sample, which allows downstream sample preparation without the risk of introducing quantification errors. During mass spectrometry analysis, the relative protein abundance between biological samples can be calculated from the intensities of heavy and light peptides. As shown by numerous applications in biological and clinical studies, SILAC represents a promising method for the elucidation of cellular and physio-pathological mechanisms and for the identification of disease biomarkers. The restriction of SILAC to the quantification of proteins from cultured cells has just been overcome with the description in a recent paper of a SILAC mouse, which will allow this technology to be applied to the differential study of -tissues and biological fluids from model animals.
基于质谱的定量蛋白质组学策略非常适合检测不同生物样品之间的整体蛋白质变化。其中，SILAC（细胞培养中氨基酸稳定同位素标记）已显示出巨大潜力。该方法极其准确，相对容易应用于从培养细胞中提取的蛋白质定量。SILAC涉及在常规培养基（“轻”蛋白质合成）或氨基酸已被其同位素标记对应物取代的培养基（“重”蛋白质合成）中进行细胞培养。待比较的细胞群体可以混合并作为单个样品处理，这使得下游样品制备不会引入定量误差。在质谱分析过程中，可以根据重肽和轻肽的强度计算生物样品之间的相对蛋白质丰度。正如在生物学和临床研究中的众多应用所示，SILAC是阐明细胞和生理病理机制以及鉴定疾病生物标志物的一种有前途的方法。最近一篇关于SILAC小鼠的论文描述克服了SILAC仅限于培养细胞蛋白质定量的限制，这将使该技术能够应用于模型动物组织和生物流体的差异研究。
