Trans-SILAC: sorting out the non-cell-autonomous proteome
Trans-SILAC: sorting out the non-cell-autonomous proteome
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DOI：
10.1038/nmeth.1513
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发表时间：
2010-11-01
期刊：
影响因子：
48
通讯作者：
Goldstein, Itamar
中科院分区：
文献类型：
--
作者：
Rechavi, Oded;Kalman, Matan;Goldstein, Itamar
非细胞自主蛋白被整合到形成紧密接触或被细菌入侵的细胞中，但识别转移蛋白的全部库一直是一个挑战。本文介绍了一种定量蛋白质组学方法，用于筛选由其他细胞或细胞内病原体合成的非细胞自主蛋白。我们的方法结合了细胞培养中氨基酸的稳定同位素标记（SILAC），高纯度细胞分选和生物信息学分析，以鉴定相关的非细胞自主蛋白库。这种“反式silac”方法使我们能够发现许多从人B细胞转移到自然杀伤细胞的蛋白质，并测量受感染的人细胞中肠沙门氏菌蛋白质的生物合成率。Trans-SILAC是检测多细胞生物不同细胞间或病原体与宿主间蛋白质交换的有效方法。
Non-cell-autonomous proteins are incorporated into cells that form tight contacts or are invaded by bacteria, but identifying the full repertoire of transferred proteins has been a challenge. Here we introduce a quantitative proteomics approach to sort out non-cell-autonomous proteins synthesized by other cells or intracellular pathogens. Our approach combines stable-isotope labeling of amino acids in cell culture (SILAC), high-purity cell sorting and bioinformatics analysis to identify the repertoire of relevant non-cell-autonomous proteins. This `trans-SILAC' method allowed us to discover many proteins transferred from human B to natural killer cells and to measure biosynthesis rates of Salmonella enterica proteins in infected human cells. Trans-SILAC should be a useful method to examine protein exchange between different cells of multicellular organisms or pathogen and host.