Allosteric control of an asymmetric transduction in a G protein-coupled receptor heterodimer
- Huazhong University of Science and Technology, China
- CNRS, INSERM, Université de Montpellier, France
- Universitat Autònoma de Barcelona, Spain
- Network Biomedical Research Center on Mental Health, Spain
- Institute for Advanced Chemistry of Catalonia (IQAC-CSIC), Spain
Figures
Figure 1 with 3 supplements
mGlu4 activates G protein in the mGlu2-4 heterodimer.
(A) Cartoons illustrating mGlu2 and mGlu4 homodimers, and mGlu2-4 (2–4 or 4–2) heterodimers with each subunit carrying the quality control C1 or C2 system as C terminal tails, and the indicated HA …
Figure 1—figure supplement 1
G protein coupling of the mGlu subunits with C1 or C2 tail.
Intracellular Ca2+ response mediated by indicated subunits co-expressed with the chimeric Gqi9 upon stimulation with DCG-IV (30 μM, green), L-AP4 (30 μM, red) or glutamate (1 mM, blue). Data are …
Figure 1—figure supplement 2
Cell surface and total expression of various mGlu dimer combination.
Quantification of cell surface-total expression of HA-tagged or Flag-tagged constructs (as indicated in Figure 1) by ELISA on intact cells transfected with indicated subunits. Data were obtained …
Figure 1—figure supplement 3
mGlu4 is responsible for G protein coupling in the mGlu2-4 heterodimer.
(A) Intracellular Ca2+ responses mediated by indicated subunits upon stimulation with increasing concentration of glutamate, in the presence of the chimeric Gqi9. (B) Cartoons illustrating the …
Figure 2 with 3 supplements
Asymmetric transduction results from the HDs in the mGlu2-4 heterodimer.
In (A), (B) and (C) cartoons illustrating the heterodimer combinations used with one or both subunits carrying the FS mutation (red cross) that prevents G protein activation are indicated on the …
Figure 2—figure supplement 1
Asymmetric transduction results from the HDs in the mGlu2-4 heterodimer.
Same as Figure 2 excepted that only the VFT is swapped, and not the entire CRD. In (A), (B) and (C) cartoons illustrating the heterodimer combinations used with one or both subunits carrying the FS …
Figure 2—figure supplement 2
Cell surface and total expression of various mGlu dimer combination.
Quantification of cell surface-total expression of HA-tagged or Flag-tagged constructs by ELISA on intact cells transfected with the indicated subunits as in Figure 2. Data were generated from the …
Figure 2—figure supplement 3
Cell surface and total expression of various mGlu dimer combination.
Quantification of cell surface-total expression of HA-tagged or Flag-tagged constructs by ELISA on intact cells transfected with the indicated subunits as in Sup Figure 4. Data were generated from …
Figure 3 with 1 supplement
Constitutive activity of disulfide-tethered mGlu2-4 heterodimer is mediated by the mGlu4 subunit.
(A) Cartoons illustrating the heterodimer combinations used with mGlu2 or mGlu4 subunits carrying the FS mutation (red cross) that prevents G protein activation (top). The red line linking both CRDs …
Figure 3—figure supplement 1
Cell surface and total expression of various mGlu dimer combination.
Quantification of cell surface-total expression of HA-tagged or Flag-tagged constructs by ELISA on intact cells transfected with the indicated subunits as in Figure 3A. Data were generated from the …
Figure 4
Allosteric regulation of mGlu4-induced signaling by mGlu2 HD.
In each panel, cartoons (color coded) illustrating the dimer compositions used are indicated on the top, and intracellular Ca2+ responses mediated by indicated dimer combinations upon stimulation …
Figure 5
Switching of the G protein coupling subunit in the mGlu2-4 heterodimer by mGlu2 PAM and mGlu4 NAM.
Intracellular Ca2+ response mediated by the indicated subunits upon stimulation with increasing concentration of glutamate with/without a mGlu4 NAM (optoGluNAM4.1, purple square, 30 μM) or a mGlu2 …
Figure 6 with 2 supplements
Asymmetric transduction by mGlu2-groupIII heterodimers.
(A–B) schemes illustrating the method used to study the coupling properties of mGlu heterodimers composed of mGlu2 (group-II) and a group-III subunit with the wild-type C-terminal tails. In (A), …
Figure 6—figure supplement 1
Expression and function of the indicated subunits.
(A) Quantification of cell expression of HA-tagged constructs by ELISA on cells transfected with the indicated subunits. (B) Intracellular Ca2+ response mediated by the indicated subunits upon …
Figure 6—figure supplement 2
Asymmetric transduction by mGlu2-groupIII heterodimers.
Same as Figure 7, except mGlu3 was used instead of mGlu2. (A) and (B) schemes illustrating the method used to study the coupling properties of mGlu heterodimers composed of mGlu3 (group-II) and a …
Figure 7
Scheme illustrating the activation mechanism and allosteric control of mGlu2-4 heterodimer.
State 1. the inactive heterodimer in its basal state. State 2: Glutamate (blue disk) activation of both subunits leads to G protein activation by mGlu4 HD, also involving a conformational change in …
Additional files
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Transparent reporting form
- https://doi.org/10.7554/eLife.26985.023
