Skip to main content
NCBI home page
British Journal of Pharmacology logoLink to British Journal of Pharmacology
. 1984 Aug;82(4):853–861. doi: 10.1111/j.1476-5381.1984.tb16482.x

Solubilization and characterization of guanine nucleotide-sensitive muscarinic agonist binding sites from rat myocardium.

C P Berrie, N J Birdsall, E C Hulme, M Keen, J M Stockton
PMCID: PMC1986921  PMID: 6478115

Abstract

Muscarinic receptors from rat myocardial membranes may be solubilized by digitonin in good yield at low temperatures in the presence of Mg2+. Under these conditions, up to 60% of the soluble receptors show high affinity binding for the potent agonist [3H]-oxotremorine-M (KA = 10(9)M-1), which is inhibited by 5'-guanylylimidodiphosphate. The muscarinic binding site labelled with [3H]-oxotremorine-M has a higher sedimentation coefficient (13.4 s) than sites labelled with a 3H antagonist in the presence of guanylylimidodiphosphate (11.6 s) and probably represents a complex between the ligand binding subunit of the receptor and a guanine nucleotide binding protein.

Full text

PDF
853

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aronstam R. S., Schuessler D. C., Jr, Eldefrawi M. E. Solubilization of muscarinic acetylcholine receptors of bovine brain. Life Sci. 1978 Oct 2;23(13):1377–1382. doi: 10.1016/0024-3205(78)90397-1. [DOI] [PubMed] [Google Scholar]
  2. Berrie C. P., Birdsall N. J., Burgen A. S., Hulme E. C. Guanine nucleotides modulate muscarinic receptor binding in the heart. Biochem Biophys Res Commun. 1979 Apr 27;87(4):1000–1005. doi: 10.1016/s0006-291x(79)80006-6. [DOI] [PubMed] [Google Scholar]
  3. Berrie C. P., Birdsall N. J., Haga K., Haga T., Hulme E. C. Hydrodynamic properties of muscarinic acetylcholine receptors solubilized from rat forebrain. Br J Pharmacol. 1984 Aug;82(4):839–851. doi: 10.1111/j.1476-5381.1984.tb16481.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bers D. M. Isolation and characterization of cardiac sarcolemma. Biochim Biophys Acta. 1979 Jul 19;555(1):131–146. doi: 10.1016/0005-2736(79)90078-6. [DOI] [PubMed] [Google Scholar]
  5. Birdsall N. J., Burgen A. S., Hulme E. C. The binding of agonists to brain muscarinic receptors. Mol Pharmacol. 1978 Sep;14(5):723–736. [PubMed] [Google Scholar]
  6. Birdsall N. J., Hulme E. C., Burgen A. The character of the muscarinic receptors in different regions of the rat brain. Proc R Soc Lond B Biol Sci. 1980 Feb 13;207(1166):1–12. doi: 10.1098/rspb.1980.0011. [DOI] [PubMed] [Google Scholar]
  7. Bokoch G. M., Katada T., Northup J. K., Hewlett E. L., Gilman A. G. Identification of the predominant substrate for ADP-ribosylation by islet activating protein. J Biol Chem. 1983 Feb 25;258(4):2072–2075. [PubMed] [Google Scholar]
  8. Carson S. Cholate-salt solubilisation of bovine brain muscarinic receptors. Biochem Pharmacol. 1982 May 1;31(9):1806–1809. doi: 10.1016/0006-2952(82)90694-3. [DOI] [PubMed] [Google Scholar]
  9. Cremo C. R., Herron G. S., Schimerlik M. I. Solubilization of the atrial muscarinic acetylcholine receptor: a new detergent system and rapid assays. Anal Biochem. 1981 Aug;115(2):331–338. doi: 10.1016/0003-2697(81)90014-2. [DOI] [PubMed] [Google Scholar]
  10. Ehlert F. J., Roeske W. R., Yamamura H. I. Regulation of muscarinic receptor binding by guanine nucleotides and N-ethylmaleimide. J Supramol Struct. 1980;14(2):149–162. doi: 10.1002/jss.400140204. [DOI] [PubMed] [Google Scholar]
  11. Gavish M., Sokolovsky M. Solubilization of muscarinic acetylcholine receptor by zwitterionic detergent from rat brain cortex. Biochem Biophys Res Commun. 1982 Dec 15;109(3):819–824. doi: 10.1016/0006-291x(82)92013-7. [DOI] [PubMed] [Google Scholar]
  12. Gorissen H., Aerts G., Laduron P. Characterization of digitonin-solubilized muscarinic receptor from rat brain. FEBS Lett. 1978 Dec 1;96(1):64–68. doi: 10.1016/0014-5793(78)81063-1. [DOI] [PubMed] [Google Scholar]
  13. Haga T., Haga K., Gilman A. G. Hydrodynamic properties of the beta-adrenergic receptor and adenylate cyclase from wild type and varient S49 lymphoma cells. J Biol Chem. 1977 Aug 25;252(16):5776–5782. [PubMed] [Google Scholar]
  14. Haga T. Molecular size of muscarinic acetylcholine receptors of rat brain. FEBS Lett. 1980 Apr 21;113(1):68–72. doi: 10.1016/0014-5793(80)80497-2. [DOI] [PubMed] [Google Scholar]
  15. Herron G. S., Miller S., Manley W. L., Schimerlik M. I. Ligand interactions with the solubilized porcine atrial muscarinic receptor. Biochemistry. 1982 Feb 2;21(3):515–520. doi: 10.1021/bi00532a016. [DOI] [PubMed] [Google Scholar]
  16. Hulme E. C., Berrie C. P., Birdsall N. J., Jameson M., Stockton J. M. Regulation of muscarinic agonist binding by cations and guanine nucleotides. Eur J Pharmacol. 1983 Oct 14;94(1-2):59–72. doi: 10.1016/0014-2999(83)90442-9. [DOI] [PubMed] [Google Scholar]
  17. Hulme E. C., Berrie C. P., Haga T., Birdsall N. J., Burgen A. S., Stockton J. Solubilisation and molecular characterisation of muscarinic acetylcholine receptors. J Recept Res. 1983;3(1-2):301–311. doi: 10.3109/10799898309041943. [DOI] [PubMed] [Google Scholar]
  18. Hurko O. Specific [3H]quinuclidinyl benzilate binding activity in digitonin-solubilized preparations from bovine brain. Arch Biochem Biophys. 1978 Oct;190(2):434–445. doi: 10.1016/0003-9861(78)90296-5. [DOI] [PubMed] [Google Scholar]
  19. Kuno T., Shirakawa O., Tanaka C. Regulation of the solubilized bovine cerebral cortex muscarinic receptor by GTP and Na+. Biochem Biophys Res Commun. 1983 May 16;112(3):948–953. doi: 10.1016/0006-291x(83)91709-6. [DOI] [PubMed] [Google Scholar]
  20. Laduron P. M., Ilien B. Solubilization of brain muscarinic, dopaminergic and serotonergic receptors: a critical analysis. Biochem Pharmacol. 1982 Jun 15;31(12):2145–2151. doi: 10.1016/0006-2952(82)90507-x. [DOI] [PubMed] [Google Scholar]
  21. Limbird L. E., Gill D. M., Lefkowitz R. J. Agonist-promoted coupling of the beta-adrenergic receptor with the guanine nucleotide regulatory protein of the adenylate cyclase system. Proc Natl Acad Sci U S A. 1980 Feb;77(2):775–779. doi: 10.1073/pnas.77.2.775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Repke H., Matthies H. Biochemical characterization of solubilized muscarinic acetylcholine receptors. Brain Res Bull. 1980 Nov-Dec;5(6):703–709. doi: 10.1016/0361-9230(80)90209-9. [DOI] [PubMed] [Google Scholar]
  23. Rodbell M. The role of hormone receptors and GTP-regulatory proteins in membrane transduction. Nature. 1980 Mar 6;284(5751):17–22. doi: 10.1038/284017a0. [DOI] [PubMed] [Google Scholar]
  24. Smith S. K., Limbird L. E. Solubilization of human platelet alpha-adrenergic receptors: evidence that agonist occupancy of the receptor stabilizes receptor--effector interactions. Proc Natl Acad Sci U S A. 1981 Jul;78(7):4026–4030. doi: 10.1073/pnas.78.7.4026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sternweis P. C., Northup J. K., Smigel M. D., Gilman A. G. The regulatory component of adenylate cyclase. Purification and properties. J Biol Chem. 1981 Nov 25;256(22):11517–11526. [PubMed] [Google Scholar]
  26. Waelbroeck M., Robberecht P., Chatelain P., Christophe J. Rat cardiac muscarinic receptors. I. Effects of guanine nucleotides on high- and low-affinity binding sites. Mol Pharmacol. 1982 May;21(3):581–588. [PubMed] [Google Scholar]
  27. Wei J. W., Sulakhe P. V. Agonist-antagonist interactions with rat atrial muscarinic cholinergic receptor sites: differential regulation by guanine nucleotides. Eur J Pharmacol. 1979 Sep 1;58(1):91–92. doi: 10.1016/0014-2999(79)90346-7. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Pharmacology are provided here courtesy of The British Pharmacological Society

RESOURCES