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. 2016 Nov 22;2:16042. doi: 10.1038/celldisc.2016.42

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Copyright © 2016 The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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C347K/R mutation transforms full-length GLP-1R into a G-protein biased receptor. (a) Position of C347 in GLP-1R. (b) Expression levels of WT and C347K/R GLP-1R detected by immunoblotting using anti-FLAG antibody for detection and β-actin antibody for normalization. G: glycosylated receptor, N: non-glycosylated receptor. ‘RG’ refers to normlalized glycosylated receptor signal relative to WT (WT set as 1.00); ‘RN’ refers to normlalized non-glycosylated receptor signal relative to WT. ‘RT’ refers to normlalized total receptor signal relative to WT. Expression of β-actin is used for normalization (see the ‘Materials and methods’ section). (c and d) EX4(1–39) dose-response curve for cAMP (c) or arrestin binding signal (d) in WT or C347K or R mutated GLP-1R. C347K/R mutant receptors do not activate arrestin signal. All values are means±s.e.m. of two independent experiments, conducted in triplicate. The activity of the receptor exposed to 1 μm EX4 was set to 100%. *: EC50 unavailable.