Direct visualization of single-molecule membrane protein interactions in living cells

by Do-Hyeon Kim, Soyeon Park, Dong-Kyun Kim, Min Gyu Jeong, Jungeun Noh, Yonghoon Kwon, Kai Zhou, Nam Ki Lee, Sung Ho Ryu

Interactions between membrane proteins are poorly understood despite their importance in cell signaling and drug development. Here, we present a co-immunoimmobilization assay (Co-II) enabling the direct observation of membrane protein interactions in single living cells that overcomes the limitations of currently prevalent proximity-based indirect methods. Using Co-II, we investigated the transient homodimerizations of epidermal growth factor receptor (EGFR) and beta-2 adrenergic receptor (β2-AR) in living cells, revealing the differential regulation of these receptors’ dimerizations by molecular conformations and microenvironment in a plasma membrane. Co-II should provide a simple, rapid, and robust platform for visualizing both weak and strong protein interactions in the plasma membrane of living cells.

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