Analysis of transient membrane protein interactions by single-molecule diffusional mobility shift assay |
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Authors: | Min Gyu Jeong Kai Zhou Soyeon Park HyeongJeon An Yonghoon Kwon Yeonho Chang Do-Hyeon Kim Sung Ho Ryu |
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Institution: | 1.Department of Life Sciences, Pohang University of Science and Technology, Pohang, Republic of Korea ;2.Department of Physics, Pohang University of Science and Technology, Pohang, Republic of Korea |
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Abstract: | 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.Subject terms: Fluorescence imaging, Super-resolution microscopy, Single-molecule biophysics |
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