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Movement of ‘gating charge’ is coupled to ligand binding in a G-protein-coupled receptor

Nature volume 444, pages 106–109 (2006)Cite this article

Abstract

Activation by agonist binding of G-protein-coupled receptors (GPCRs) controls most signal transduction processes1. Although these receptors span the cell membrane, they are not considered to be voltage sensitive. Recently it was shown that both the activity of GPCRs2,3,4,5 and their affinity towards agonists6 are regulated by membrane potential. However, it remains unclear whether GPCRs intrinsically respond to changes in membrane potential. Here we show that two prototypical GPCRs, the m2 and m1 muscarinic receptors (m2R and m1R), display charge-movement-associated currents analogous to ‘gating currents’ of voltage-gated channels. The gating charge–voltage relationship of m2R correlates well with the voltage dependence of the affinity of the receptor for acetylcholine. The loop that couples m2R and m1R to their G protein has a crucial function in coupling voltage sensing to agonist-binding affinity. Our data strongly indicate that GPCRs serve as sensors for both transmembrane potential and external chemical signals.

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Figure 1: Gating currents in m2R-expressing oocytes.
Figure 2: Voltage dependences of gating currents of m1R and m2R, and of the agonist affinity of m2R.
Figure 3: Gating currents and dose–response curves of mutated m2R.

References

  1. Gether, U. Uncovering molecular mechanisms involved in activation of G protein- coupled receptors. Endocr. Rev. 21, 90–113 (2000)

    Article  CAS  Google Scholar 

  2. Marty, A. & Tan, Y. P. The initiation of calcium release following muscarinic stimulation in rat lacrimal glands. J. Physiol. (Lond.) 419, 665–687 (1989)

    Article  CAS  Google Scholar 

  3. Ong, B. H. et al. G protein modulation of voltage-sensitive muscarinic receptor signaling in mouse pancreatic acinar cells. Pflügers Arch. 441, 604–610 (2001)

    Article  CAS  Google Scholar 

  4. Martinez-Pinna, J., Tolhurst, G., Gurung, I. S., Vandenberg, J. I. & Mahaut-Smith, M. P. Sensitivity limits for voltage control of P2Y receptor-evoked Ca2+ mobilization in the rat megakaryocyte. J. Physiol. (Lond.) 555, 61–70 (2004)

    Article  CAS  Google Scholar 

  5. Martinez-Pinna, J. et al. Direct voltage control of signaling via P2Y1 and other Galphaq-coupled receptors. J. Biol. Chem. 280, 1490–1498 (2005)

    Article  CAS  Google Scholar 

  6. Ben-Chaim, Y., Tour, O., Dascal, N., Parnas, I. & Parnas, H. The M2 muscarinic G-protein-coupled receptor is voltage-sensitive. J. Biol. Chem. 278, 22482–22491 (2003)

    Article  CAS  Google Scholar 

  7. Hodgkin, A. L. & Huxley, A. F. A quantitative description of membrane current and its application to conduction and excitation in nerve. J. Physiol. (Lond.) 117, 500–544 (1952)

    Article  CAS  Google Scholar 

  8. Armstrong, C. M. & Bezanilla, F. Currents related to movement of the gating particles of the sodium channels. Nature 242, 459–461 (1973)

    Article  ADS  CAS  Google Scholar 

  9. Rakowski, R. F. et al. Charge translocation by the Na/K pump. Ann. NY Acad. Sci. 834, 231–243 (1997)

    Article  ADS  CAS  Google Scholar 

  10. Loo, D. D., Hazama, A., Supplisson, S., Turk, E. & Wright, E. M. Relaxation kinetics of the Na+/glucose cotransporter. Proc. Natl Acad. Sci. USA 90, 5767–5771 (1993)

    Article  ADS  CAS  Google Scholar 

  11. Murata, Y., Iwasaki, H., Sasaki, M., Inaba, K. & Okamura, Y. Phosphoinositide phosphatase activity coupled to an intrinsic voltage sensor. Nature 435, 1239–1243 (2005)

    Article  ADS  CAS  Google Scholar 

  12. Stefani, E. & Bezanilla, F. Cut-open oocyte voltage-clamp technique. Methods Enzymol. 293, 300–318 (1998)

    Article  CAS  Google Scholar 

  13. Bezanilla, F. & Armstrong, C. M. Inactivation of the sodium channel. I. Sodium current experiments. J. Gen. Physiol. 70, 549–566 (1977)

    Article  CAS  Google Scholar 

  14. Haga, K., Haga, T. & Ichiyama, A. Reconstitution of the muscarinic acetylcholine receptor. Guanine nucleotide-sensitive high affinity binding of agonists to purified muscarinic receptors reconstituted with GTP-binding proteins (Gi and Go). J. Biol. Chem. 261, 10133–10140 (1986)

    CAS  Google Scholar 

  15. Cohen-Armon, M. & Sokolovsky, M. Evidence for involvement of the voltage-dependent Na+ channel gating in depolarization-induced activation of G-proteins. J. Biol. Chem. 268, 9824–9838 (1993)

    CAS  Google Scholar 

  16. Ilouz, N., Branski, L., Parnis, J., Parnas, H. & Linial, M. Depolarization affects the binding properties of muscarinic acetylcholine receptors and their interaction with proteins of the exocytic apparatus. J. Biol. Chem. 274, 29519–29528 (1999)

    Article  CAS  Google Scholar 

  17. Kenakin, T. Principles: receptor theory in pharmacology. Trends Pharmacol. Sci. 25, 186–192 (2004)

    Article  CAS  Google Scholar 

  18. De Lean, A., Stadel, J. M. & Lefkowitz, R. J. A ternary complex model explains the agonist-specific binding properties of the adenylate cyclase-coupled β-adrenergic receptor. J. Biol. Chem. 255, 7108–7117 (1980)

    CAS  Google Scholar 

  19. Lefkowitz, R. J., Cotecchia, S., Samama, P. & Costa, T. Constitutive activity of receptors coupled to guanine nucleotide regulatory proteins. Trends Pharmacol. Sci. 14, 303–307 (1993)

    Article  CAS  Google Scholar 

  20. Lechleiter, J. et al. Distinct sequence elements control the specificity of G protein activation by muscarinic acetylcholine receptor subtypes. EMBO J. 9, 4381–4390 (1990)

    Article  CAS  Google Scholar 

  21. Wess, J. G-protein-coupled receptors: molecular mechanisms involved in receptor activation and selectivity of G-protein recognition. FASEB J. 11, 346–354 (1997)

    Article  CAS  Google Scholar 

  22. Wess, J., Bonner, T. I., Dorje, F. & Brann, M. R. Delineation of muscarinic receptor domains conferring selectivity of coupling to guanine nucleotide-binding proteins and second messengers. Mol. Pharmacol. 38, 517–523 (1990)

    CAS  Google Scholar 

  23. Kubo, T. et al. Location of a region of the muscarinic acetylcholine receptor involved in selective effector coupling. FEBS Lett. 241, 119–125 (1988)

    Article  CAS  Google Scholar 

  24. Zhu, S. Z., Wang, S. Z., Hu, J. & el-Fakahany, E. E. An arginine residue conserved in most G protein-coupled receptors is essential for the function of the m1 muscarinic receptor. Mol. Pharmacol. 45, 517–523 (1994)

    CAS  Google Scholar 

  25. Hulme, E. C., Curtis, C. A., Page, K. M. & Jones, P. G. The role of charge interactions in muscarinic agonist binding, and receptor-response coupling. Life Sci. 56, 891–898 (1995)

    Article  CAS  Google Scholar 

  26. Ohana, L., Barchad, O., Parnas, I. & Parnas, H. The metabotropic glutamate G-protein-coupled receptors mGluR3 and mGluR1a are voltage sensitive. J. Biol. Chem. 281, 24204–24215 (2006)

    Article  CAS  Google Scholar 

  27. Linden, D. J. Long-term synaptic depression in the mammalian brain. Neuron 12, 457–472 (1994)

    Article  CAS  Google Scholar 

  28. Bradley, S. R. et al. Activation of group II metabotropic glutamate receptors inhibits synaptic excitation of the substantia nigra pars peticulata. J. Neurosci. 20, 3085–3094 (2000)

    Article  CAS  Google Scholar 

  29. Parnas, H., Segel, L., Dudel, J. & Parnas, I. Autoreceptors, membrane potential and the regulation of transmitter release. Trends Neurosci. 23, 60–68 (2000)

    Article  CAS  Google Scholar 

  30. Peleg, S., Varon, D., Ivanina, T., Dessauer, C. W. & Dascal, N. G. αi controls the gating of the G protein-activated K+ channel, GIRK. Neuron 33, 87–99 (2002)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank T. Kubo for providing us with the clones for the m2R/L3-m1 and m1R/L3-m2 constructs. This work was supported by a grant to J. Dudel, I.P. and H.P. from the Deutsche Forschungsgemeinschaft, and by NIH grants to N.D., B.C. and to F.B.

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Author notes

  1. Yair Ben-Chaim

    Present address: Neuroscience Department, The John Hopkins University Medical School, Baltimore, MD21205, USA

Authors and Affiliations

  1. Department of Neurobiology, The Hebrew University, Jerusalem, 91904, Israel

    Yair Ben-Chaim, Itzchak Parnas & Hanna Parnas

  2. Department of Physiology, University of Wisconsin, Madison, Wisconsin, 53706, USA

    Baron Chanda

  3. Department of Physiology and Pharmacology, Tel Aviv University, Tel-Aviv, 69978, Israel

    Nathan Dascal

  4. Institute for Molecular Pediatric Sciences, University of Chicago, Chicago, Illinois, 60637, USA

    Francisco Bezanilla

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Correspondence to Hanna Parnas.

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Ben-Chaim, Y., Chanda, B., Dascal, N. et al. Movement of ‘gating charge’ is coupled to ligand binding in a G-protein-coupled receptor. Nature 444, 106–109 (2006). https://doi.org/10.1038/nature05259

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