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KMID : 0620920210530020291
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Experimental & Molecular Medicine
2021 Volume.53 No. 2 p.291 ~ p.299
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Analysis of transient membrane protein interactions by single-molecule diffusional mobility shift assay
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Jeong Min-Gyu
Zhou Kai
Park So-Yeon
An Hyeong-Jeon
Kwon Yong-Hoon
Chang Yeon-Ho
Kim Do-Hyeon
Ryu Sung-Ho
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Abstract
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Various repertoires of membrane protein interactions determine cellular responses to diverse environments around cells dynamically in space and time. Current assays, however, have limitations in unraveling these interactions in the physiological states in a living cell due to the lack of capability to probe the transient nature of these interactions on the crowded membrane. Here, we present a simple and robust assay that enables the investigation of transient protein interactions in living cells by using the single-molecule diffusional mobility shift assay (smDIMSA). Utilizing smDIMSA, we uncovered the interaction profile of EGFR with various membrane proteins and demonstrated the promiscuity of these interactions depending on the cancer cell line. The transient interaction profile obtained by smDIMSA will provide critical information to comprehend the crosstalk among various receptors on the plasma membrane.
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KEYWORD
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Fluorescence imaging, Single-molecule biophysics, Super-resolution microscopy
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