Quantification of extracellular proteins, protein complexes and mRNAs in single cells by proximity sequencing
Quantification of extracellular proteins, protein complexes and mRNAs in single cells by proximity sequencing
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DOI：
10.1038/s41592-022-01684-z
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发表时间：
2022-12-01
期刊：
影响因子：
48
通讯作者：
Tay，Savas
中科院分区：
文献类型：
--
作者：
Vistain，Luke;Phan，Hoang Van;Tay，Savas
我们提出近距离测序（Prox-seq），用于同时测量数千个单细胞中的蛋白质，蛋白质复合物和mrna。Prox-seq结合了接近连接法和单细胞测序，从目标蛋白的所有成对组合中测量蛋白质及其复合物，提供二次缩放的多路复用。我们验证了Prox-seq，并分析了T细胞和B细胞的混合物，表明它能准确地识别这些细胞类型，并检测出众所周知的蛋白质复合物。接下来，通过研究人外周血单核细胞，我们发现naïve CD8+T细胞显示蛋白复合物CD8 - cd9。最后，我们研究了巨噬细胞中toll样受体（TLR）信号传导过程中的蛋白质相互作用。我们观察了信号特异性蛋白复合物的形成，发现了CD36共受体在脂多糖（TLR4）和Pam2CSK4 （TLR2）刺激下的活性和附加信号整合，并表明蛋白复合物的量化识别了巨噬细胞接受的信号输入。Prox-seq提供了一种未开发的单细胞表型测量方式，可以发现不同细胞类型中未表征的蛋白质相互作用。
We present proximity sequencing (Prox-seq) for simultaneous measurement of proteins, protein complexes and mRNAs in thousands of single cells. Prox-seq combines proximity ligation assay with single-cell sequencing to measure proteins and their complexes from all pairwise combinations of targeted proteins, providing quadratically scaled multiplexing. We validate Prox-seq and analyze a mixture of T cells and B cells to show that it accurately identifies these cell types and detects well-known protein complexes. Next, by studying human peripheral blood mononuclear cells, we discover that naïve CD8+T cells display the protein complex CD8–CD9. Finally, we study protein interactions during Toll-like receptor (TLR) signaling in human macrophages. We observe the formation of signal-specific protein complexes, find CD36 co-receptor activity and additive signal integration under lipopolysaccharide (TLR4) and Pam2CSK4 (TLR2) stimulation, and show that quantification of protein complexes identifies signaling inputs received by macrophages. Prox-seq provides access to an untapped measurement modality for single-cell phenotyping and can discover uncharacterized protein interactions in different cell types.