Shenoy Anjana, Geiger Tamar
Department of Human Molecular Genetics and Biochemistry, Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, 69978, Israel.
Expert Rev Proteomics. 2015 Feb;12(1):13-9. doi: 10.1586/14789450.2015.982538. Epub 2014 Nov 18.
Stable isotope labeling with amino acids in cell culture (SILAC) has risen as a powerful quantification technique in mass spectrometry (MS)-based proteomics in classical and modified forms. Previously, SILAC was limited to cultured cells because of the requirement of active protein synthesis; however, in recent years, it was expanded to model organisms and tissue samples. Specifically, the super-SILAC technique uses a mixture of SILAC-labeled cells as a spike-in standard for accurate quantification of unlabeled samples, thereby enabling quantification of human tissue samples. Here, we highlight the recent developments in super-SILAC and its application to the study of clinical samples, secretomes, post-translational modifications and organelle proteomes. Finally, we propose super-SILAC as a robust and accurate method that can be commercialized and applied to basic and clinical research.
细胞培养中氨基酸稳定同位素标记(SILAC)已成为基于质谱(MS)的蛋白质组学中一种强大的定量技术,有经典和改良等形式。以前,由于需要活跃的蛋白质合成,SILAC仅限于培养细胞;然而,近年来,它已扩展到模式生物和组织样本。具体而言,超级SILAC技术使用SILAC标记细胞的混合物作为内标,用于准确量化未标记样本,从而能够对人体组织样本进行定量分析。在此,我们重点介绍超级SILAC的最新进展及其在临床样本、分泌蛋白质组、翻译后修饰和细胞器蛋白质组研究中的应用。最后,我们提出超级SILAC是一种稳健且准确的方法,可实现商业化并应用于基础研究和临床研究。