Department of Immunology, Weizmann Institute of Science, Rehovot, 76100, Israel.
Department of Biological Services, Weizmann Institute of Science, Rehovot, 76100, Israel.
Nat Commun. 2017 Nov 15;8(1):1524. doi: 10.1038/s41467-017-01808-6.
In spite of recent advances in proteomics, quantitative analyses of protein-protein interactions (PPIs) or post-translational modifications (PTMs) in rare cell populations remain challenging. This is in particular true for analyses of rare immune and/or stem cell populations that are directly isolated from humans or animal models, and which are often characterized by multiple surface markers. To overcome these limitations, here we have developed proximity ligation imaging cytometry (PLIC), a protocol for proteomic analysis of rare cells. Specifically, by employing PLIC on medullary thymic epithelial cells (mTECs), which serve as a paradigm for a rare immune population, we demonstrate that PLIC overcomes the inherent limitations of conventional proteomic approaches and enables a high-resolution detection and quantification of PPIs and PTMs at a single cell level.
尽管蛋白质组学领域最近取得了一些进展，但在稀有细胞群体中定量分析蛋白质-蛋白质相互作用（PPIs）或翻译后修饰（PTMs）仍然具有挑战性。对于直接从人类或动物模型中分离出来的稀有免疫细胞和/或干细胞群体的分析尤其如此，这些细胞群体通常具有多个表面标志物。为了克服这些限制，我们在这里开发了邻近连接成像细胞计量术（PLIC），这是一种用于稀有细胞的蛋白质组学分析的方案。具体来说，通过在作为稀有免疫群体范例的骨髓胸腺上皮细胞（mTECs）上使用 PLIC，我们证明了 PLIC 克服了传统蛋白质组学方法的固有局限性，并能够在单细胞水平上高分辨率地检测和定量 PPI 和 PTM。
