The coupling of the M2 muscarinic receptor to its G protein is voltage dependent

Autoři: Yair Ben-Chaim aff001;  Chava Broide aff002;  Hanna Parnas aff002
Působiště autorů: Department of Natural and Life Sciences, The Open University of Israel, Ra'anana, Israel aff001;  Department of Neurobiology, Institute of Life Sciences, The Hebrew University, Jerusalem, Israel aff002
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article


G protein coupled receptors (GPCRs) participate in the majority of signal transduction processes in the body. Specifically, the binding of an external agonist promotes coupling of the GPCR to its G protein and this, in turn, induces downstream signaling. Recently, it was shown that agonist binding to the M2 muscarinic receptor (M2R) and to other GPCRs is voltage dependent. Here we examine, whether the coupling of the M2R to its G protein is also voltage-dependent. We first show, in Xenopus oocytes, that the activity of the M2R in the absence of agonist (constitutive activity) can be used to report the coupling. We then show that the coupling is, by itself, voltage dependent. This novel finding is of physiological importance, as it shows that the actual signal transduction, whose first step is the coupling of the GPCR to its cognate G protein, is voltage dependent.

Klíčová slova:

Depolarization – G protein coupled receptors – Membrane potential – Oocytes – Signal transduction – Xenopus oocytes – Atropine – G-protein signaling


1. Gether U. Uncovering molecular mechanisms involved in activation of G protein- coupled receptors. Endocr Rev. 2000;21: 90–113. doi: 10.1210/edrv.21.1.0390 10696571

2. Ben-Chaim Y, Chanda B, Dascal N, Bezanilla F, Parnas I, Parnas H. Movement of “gating charge” is coupled to ligand binding in a G-protein-coupled receptor. Nature. 2006;444: 106–109. Available: doi: 10.1038/nature05259 17065983

3. Ben-Chaim Y, Tour O, Dascal N, Parnas I, Parnas H. The M2 muscarinic G-protein-coupled receptor is voltage-sensitive. J Biol Chem. 2003;278: 22482–22491. Available: doi: 10.1074/jbc.M301146200 12684524

4. Ohana L, Barchad O, Parnas I, Parnas H. The metabotropic glutamate G-protein-coupled receptors mGluR3 and mGluR1a are voltage sensitive. J Biol Chem. 2006. Available:

5. Sahlholm K, Nilsson J, Marcellino D, Fuxe K, Århem P. Voltage-dependence of the human dopamine D2 receptor. Synapse. 2008;62: 476–480. doi: 10.1002/syn.20509 18361445

6. Rinne A, Birk A, Bünemann M. Voltage regulates adrenergic receptor function. Proc Natl Acad Sci. 2013;110: 1536 LP– 1541. Available:

7. Mahaut-Smith MP, Martinez-Pinna J, Gurung IS. A role for membrane potential in regulating GPCRs? Trends Pharmacol Sci. 2008;29: 421–429. Available: doi: 10.1016/ 18621424

8. Navarro-Polanco RA, Galindo EGM, Ferrer-Villada T, Arias M, Rigby JR, Sánchez-Chapula JA, et al. Conformational changes in the M2 muscarinic receptor induced by membrane voltage and agonist binding. J Physiol. 2011;589: 1741–1753. doi: 10.1113/jphysiol.2010.204107 21282291

9. Birk A, Rinne A, Bünemann M. Membrane Potential Controls the Efficacy of Catecholamine-induced β1-Adrenoceptor Activity. J Biol Chem. 2015/09/25. 2015;290: 27311–27320. doi: 10.1074/jbc.M115.665000 26408198

10. Sahlholm K, Barchad-Avitzur O, Marcellino D, Gómez-Soler M M, Fuxe K, Ciruela F, et al. Agonist-specific voltage sensitivity at the dopamine D2S receptor- Molecular determinants and relevance to therapeutic ligands. Neuropharmacology. 2011;61: 937–949. Available: doi: 10.1016/j.neuropharm.2011.06.022 21752340

11. Ben Chaim Y, Bochnik S, Parnas I, Parnas H. Voltage Affects the Dissociation Rate Constant of the m2 Muscarinic Receptor. PLoS One. 2013;8: e74354. Available: doi: 10.1371/journal.pone.0074354 24019965

12. Nelson CP. Constitutive Activity and Inverse Agonism at the M2 Muscarinic Acetylcholine Receptor. J Pharmacol Exp Ther. 2005;316: 279–288. doi: 10.1124/jpet.105.094383 16188951

13. Jan J, Lucie B, E. E-FE, Stanislav T. Constitutive activity of the M1–M4 subtypes of muscarinic receptors in transfected CHO cells and of muscarinic receptors in the heart cells revealed by negative antagonists. FEBS Lett. 2000;377: 275–279. doi: 10.1016/0014-5793(95)01360-1

14. Spalding TA, Burstein ES. Constitutive Activity of Muscarinic Acetylcholine Receptors. J Recept Signal Transduct. 2006;26: 61–85. doi: 10.1080/10799890600567349 16595339

15. Lefkowitz RJ, Cotecchia S, Samama P, Costa T. Constitutive activity of receptors coupled to guanine nucleotide regulatory proteins. Trends Pharmacol Sci. 1993;14: 303–307. doi: 10.1016/0165-6147(93)90048-O 8249148

16. Peleg S, Varon D, Ivanina T, Dessauer CW, Dascal N. G(alpha)(i) controls the gating of the G protein-activated K(+) channel, GIRK. Neuron. 2002;33: 87–99. Available: doi: 10.1016/s0896-6273(01)00567-0 11779482

17. Vorobiov D, Bera AK, Keren-Raifman T, Barzilai R, Dascal N. Coupling of the muscarinic m2 receptor to G protein-activated K(+) channels via Galpha(z) and a receptor-Galpha(z) fusion protein. Fusion between the receptor and Galpha(z) eliminates catalytic (collision) coupling. J Biol Chem. 2000;275: 4166–4170. Available: doi: 10.1074/jbc.275.6.4166 10660578

18. Barchad-Avitzur O, Priest MF, Dekel N, Bezanilla F, Parnas H, Ben-Chaim Y. A Novel Voltage Sensor in the Orthosteric Binding Site of the M2 Muscarinic Receptor. Biophys J. 2016;111: 1396–1408. doi: 10.1016/j.bpj.2016.08.035 27705763

19. Dascal N. Signalling via the G protein-activated K+ channels. Cell Signal. 1997;9: 551–573. Available: doi: 10.1016/s0898-6568(97)00095-8 9429760

20. Sahlholm K, Marcellino D, Nilsson J, Fuxe K, Ãrhem P. Differential voltage-sensitivity of D2-like dopamine receptors. Biochem Biophys Res Commun. 2008;374: 496–501. Available: doi: 10.1016/j.bbrc.2008.07.052 18652803

Článek vyšel v časopise


2019 Číslo 10
Nejčtenější tento týden