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Ion-Channel-Coupled Receptor-Based Platform for a Real-Time Measurement of G-Protein-Coupled Receptor Activities

Cited 11 time in Web of Science Cited 13 time in Scopus
Authors
Lim, Jong Hyun; Oh, Eun Hae; Park, Juhun; Hong, Seunghun; Park, Tai Hyun
Issue Date
2015
Publisher
American Chemical Society
Citation
ACS Nano, vol.9 no.2, pp. 1699-1706
Keywords
ion-channel-coupled receptornanovesiclecarbon nanotubeKir6.2G-protein-coupled receptor
Abstract
A simple but efficient measurement platform based on ion-channelcoupled receptors and nanovesicles was developed for monitoring the real-time activity of G-protein-coupled receptors (GPCRs). In this work, an olfactory receptor (OR), the most common class A GPCR, was covalently fused with a Kir6.2 channel so that the GPCR action directly induced the opening of the ion channels and changes in the electrical membrane potential without complex cellular signaling processes. This strategy reduced the measurement errors caused by instability of various cellular components. In addition, rather than using whole cells, a cell-surface-derived nanovesicle was used to preserve the membrane-integrated structure of GPCRs and to exclude case-dependent cellular conditions. Another merit of using the nanovesicle is that nanovesicles can be easily combined with nanomaterial-based field-effect transistors (FETs) to build a sensitive and stable measurement platform to monitor GPCR activities with high sensitivity in real-time. Using a platform based on carbon nanotube FETs and nanovesicles carrying Kir6.2-channel-coupled ORs, we monitored the real-time response of ORs to their ligand molecules. Significantly, since this platform does not rely on rather unstable cell signaling pathways, our platform could be utilized for a rather long time period without losing its functionality. This system can be utilized extensively for simple and sensitive analysis of the activities of various GPCRs and should enable various academic and practical applications.
URI
http://hdl.handle.net/10371/95464
DOI
https://doi.org/10.1021/nn506494e
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College of Engineering/Engineering Practice School (공과대학/대학원)Dept. of Chemical and Biological Engineering (화학생물공학부)Journal Papers (저널논문_화학생물공학부)
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