The C-Terminus of the mu Opioid Receptor Is Critical in G-Protein Interaction as Demonstrated by a Novel Graphene Biosensor
Publication in refereed journal


摘要Several water-soluble variants of the human mu opioid receptor (wsMORs) have been designed and expressed, which enables the detection of opioids in the nM to pM range using biosensing platforms. The tools previously developed allowed us to investigate MOR and G-protein interactions in a lipid free system to demonstrate that the lipid bilayer might not be essential for the G-protein recognition and binding. In this study, we are able to investigate G-protein interactions with MOR by using graphene enabled technology, in a lipid free system, with a high sensitivity in a real time manner. A new wsMOR with the native C-terminus was designed, expressed and then immobilized on the surfaces of scalable graphene field effect transistor (GFET)-based biosensors, enabling the recording of wsMOR/G-protein interaction with an electronic readout. G-protein only interacts with the wsMOR in the presence of the native MOR C-terminus with a KA of 32.3± 11.1 pM. The electronic readout of such interaction is highly reproducible with little variance across 50 devices in one biosensor array. For devices with receptors that do not have the native C-terminus, no significant electronic response was observed in the presence of G-protein, indicating an absence of interaction. These findings reveal that lipid environment is not essential for the G-protein interaction with MOR, however, the C-terminus of MOR is essential for G-protein recognition and high affinity binding. A system to detect MOR-G protein interaction is developed. wsMOR-G2_Cter provides a novel tool to investigate the role of C terminus in the signaling pathway.
著者Wen C., Selling B., Yeliseev A., Xi J., Perez-Aguilar J.M., Gao Z., Saven J.G., Charlie Johnson A.T., Liu R.
期刊名稱IEEE Sensors Journal
出版社Institute of Electrical and Electronics Engineers Inc.
頁次5758 - 5762
關鍵詞Biosensors, C-terminus, G-protein, graphene, mu opioid receptor, water-soluble

上次更新時間 2021-14-10 於 23:51