Department of Life Sciences, Pohang University of Science and Technology, Pohang, Republic of Korea.
School of Interdisciplinary Bioscience and Bioengineering, Pohang University of Science and Technology, Pohang, Republic of Korea.
PLoS Biol. 2018 Dec 13;16(12):e2006660. doi: 10.1371/journal.pbio.2006660. eCollection 2018 Dec.
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.
尽管膜蛋白在细胞信号转导和药物开发中具有重要作用，但它们之间的相互作用仍未被充分了解。在这里，我们提出了一种共免疫固定化测定法（Co-II），该方法可直接观察单个活细胞中膜蛋白的相互作用，克服了目前流行的基于邻近性的间接方法的局限性。使用 Co-II，我们研究了表皮生长因子受体（EGFR）和β2 肾上腺素能受体（β2-AR）在活细胞中的瞬时同源二聚化，揭示了这些受体的二聚化在质膜中受到分子构象和微环境的差异调节。Co-II 应该为可视化活细胞质膜中弱和强蛋白相互作用提供一个简单、快速和强大的平台。
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