小柯机器人

美国芝加哥大学Sava Tay研究团队通过邻近测序对单细胞中的细胞外蛋白、蛋白复合物和mRNA进行定量分析。2022年12月1日，《自然—方法学》杂志在线发表了这项成果。

研究人员提出了临近测序（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将接近结扎测定与单细胞测序结合起来，从所有目标蛋白质的成对组合中测量蛋白质及其复合物，且提供了二次方规模的复用。研究人员验证了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，并检测著名的蛋白质复合物。接下来，通过研究人类外周血单核细胞，研究人员发现新生的CD8⁺T细胞显示了CD8-CD9的蛋白复合物。

最后，研究人员揭示了人类巨噬细胞中Toll样受体（TLR）信号传导过程中的蛋白质相互作用。研究人员观察了信号特异性蛋白复合物的形成，并发现在脂多糖（TLR4）和Pam2CSK4（TLR2）的刺激下，CD36共受体的活性和附加信号的整合，并显示蛋白复合物的量化可以确定巨噬细胞接收的信号输入。Prox-seq为单细胞表型提供了一个未开发的测量模式，并能发现不同类型细胞中未定性的蛋白质相互作用。

附：英文原文

Title: Quantification of extracellular proteins, protein complexes and mRNAs in single cells by proximity sequencing

Author: Vistain, Luke, Van Phan, Hoang, Keisham, Bijentimala, Jordi, Christian, Chen, Mengjie, Reddy, Sai T., Tay, Sava

Issue&Volume: 2022-12-01

Abstract: 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 nave 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.

DOI: 10.1038/s41592-022-01684-z

Source: https://www.nature.com/articles/s41592-022-01684-z

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex