Abstract
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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Cell Line
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Cell Membrane / metabolism
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ErbB Receptors / physiology
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Humans
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Immunoprecipitation / methods*
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Membrane Proteins / physiology
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Protein Binding / physiology
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Protein Interaction Mapping / methods*
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Receptors, Adrenergic, beta-2 / physiology
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Signal Transduction
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Single-Cell Analysis / methods*
Substances
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Membrane Proteins
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Receptors, Adrenergic, beta-2
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ErbB Receptors
Grants and funding
National Research Foundation of Korea (grant number NRF-2016K1A1A2912722). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. National Research Foundation of Korea (grant number NRF-2016R1A6A3A11935984). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Samsung Science and Technology Foundation (grant number SSTF-BA1402-11). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.