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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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The interaction interface between ECD and TMD localizes to the ECL of GLP-1R. Top left: cartoon representation of receptor TMD co-expressed with membrane-tethered EX4-ECD fusion (EX4-ECD-M); this construct has been used in a, c and e. Top right: cartoon representation of the hormone-fused receptor; this construct has been used in b, d and f. (a, c and e) cAMP signal produced by EX4-ECD-M and GLP-1R TMD with indicated mutations in ECL1, 2 and 3. (b, d and f) cAMP signal produced by EX4-5xGSA-GLP-1R FL construct harboring mutations in the same sites as the constructs in (a, c and e). All signal are indicated as relative to control (EX4-5xGSA-GLP-1R FL WT, labeled as C in a, c and e, as C with bracket in b, d and f), which was set to 100%. EX4-ECD-M: membrane-tethered EX4-ECD fusion. Error bars=s.d., all experiments were performed as three independent transfection experiments; two-tailed Student’s t test was used to determine P-values for data points versus WT: NS>0.05; *⩽0.05; **⩽0.01; ***⩽0.001; ****⩽0.0001. See Supplementary Figure S3B–D for activities normalized to expression levels.