Kalxdorf Mathias, Eberl Hans Christian, Bantscheff Marcus
Cellzome, A GSK Company, Meyerhofstrasse 1, 69117, Heidelberg, Germany.
Methods Mol Biol. 2017;1647:47-59. doi: 10.1007/978-1-4939-7201-2_3.
The analysis of the cell surface accessible proteome provides invaluable information about cellular identity, cellular functions, and interactions. Cell surface labeling in combination with quantitative proteomics enables the unbiased identification and quantification of cell surface proteins. We describe a fast, efficient, and robust protocol for the enrichment of the N-linked plasma membrane glycoproteome and subsequent analysis by mass spectrometry. Precise and multiplexed quantification of relative changes of cell surface protein presentation is enabled by an isobaric labeling strategy.
细胞表面可及蛋白质组的分析为细胞身份、细胞功能及相互作用提供了极有价值的信息。细胞表面标记与定量蛋白质组学相结合,能够对细胞表面蛋白进行无偏倚的鉴定和定量。我们描述了一种快速、高效且稳健的方案,用于富集N-连接的质膜糖蛋白质组,并随后通过质谱进行分析。同量异位标记策略能够实现细胞表面蛋白表达相对变化的精确和多重定量。