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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Ligand Binding Properties of Muscarinic Acetylcholine Receptors in Caenorhabditis elegans

  • You, Suck-Jong (Department of Biochemistry, College of Natural Sciences, Chungbuk National University) ;
  • Choi, Jung-Do (Department of Biochemistry, College of Natural Sciences, Chungbuk National University) ;
  • Cho, Nam-Jeong (Department of Biochemistry, College of Natural Sciences, Chungbuk National University)
  • Received : 1996.07.24
  • Published : 1996.11.30
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Abstract

Ligand binding properties of muscarinic acetylcholine receptors (mAChRs) in the nematode Caenorhabditis elegans (C. elegans) were characterized by using filtration binding assays. Scatchard analysis using $[^{3}H]N-methylscopolamine$ ($[^{3}H]NMS$) showed that the dissociation constant ($K_d$) and the maximum binding value ($B_{max}$) were $3.3{\pm}0.8{\times}10^{10}$ M and $9.0{\pm}1.1$ fmol/mg protein, respectively. Binding competition experiments indicated that the affinities of C. elegans mAChRs to atropine, scopolamine, and oxotremorine were similar to those of mammalian mAChRs. Pirenzepine binding experiments revealed that the binding pattern of mAChRs in C. elegans closely resembled that of mAChRs in rat brain, suggesting that the receptors consist primarily of Ml subtype. The affinity of mAChRs for oxotrernorine was significantly affected by guanylylimidodiphosphate (Gpp(NH)p), a non hydrolyzable GTP analog, suggesting that mAChRs in C. elegans might be coupled to G proteins. The data presented here indicate the possibility that C. elegans provides a living animal model to study the action mode of the muscarinic cholinergic system.

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References

  1. Mol. Pharmacol. v.44 Blake, A.D.;Anthony, N.M.;Chen, H.H.;Harrison, J.B.;Nathanson, N.M.;Sattelle, D.B.
  2. Science v.237 Bonner, T.I.;Buckley, N.J.;Young, A.C.;Brann, M.R. https://doi.org/10.1126/science.3037705
  3. Science v.263 Chalfie, M.;Tu, Y.;Euskirchen, G.;Ward. W.W.;Prasher, D.C. https://doi.org/10.1126/science.8303295
  4. Biochem. Pharmacol. v.22 Cheng, Y.C.;Prusoff, W.H. https://doi.org/10.1016/0006-2952(73)90196-2
  5. Proc. Natl. Acad. Sci. USA v.85 Conklin, B.R.;Brann, M.R.;Buckley, N.J.;Ma, A.L.;Bonner, T.I.;Axelrod, J. https://doi.org/10.1073/pnas.85.22.8698
  6. J. Neurosci. v.3 Culotti, J.G.;Klein, W.L. https://doi.org/10.1523/JNEUROSCI.03-02-00359.1983
  7. Annu. Rev. Biochem. v.60 Dohlman, H.G.;Thorner, J.;Caron, M.G.;Lefkowitz, R.J. https://doi.org/10.1146/annurev.bi.60.070191.003253
  8. J. Pharmacol. Exp. Ther. v.242 Doods, H.N.;Mathy, M.J.;Davidesko, D.;Van Charldorp, K.J.;De Jonge, A.;Van Zwieten, P.A.
  9. Proc. Natl. Acad. Sci. USA v.88 Gutkind, J.S.;Novotny, E.A.;Brann, M.R.;Robbins, K.C. https://doi.org/10.1073/pnas.88.11.4703
  10. Nature v.283 Hammer, R.;Berrie, C.P.;Birdsall, N.J.M.;Burgen, A.S.V.;Hulme, E.C. https://doi.org/10.1038/283090a0
  11. Science v.240 Kenyon, C. https://doi.org/10.1126/science.3287621
  12. Nature v.323 Kubo, T.;Fukuda, K.;Mikami, A.;Maeda, A.;Takahashi, H.;Mishina, M.;Haga, T.;Haga, K.;lchiyama, A.;Kangawa, K.;Kojima, M.;Matsuo, H.;Hirose, T.;Numa, S. https://doi.org/10.1038/323411a0
  13. J. Biol. Chem. v.193 Lowry, O.H.;Rosebrough, N.J.;Farr, A.L.;Randall, R.J.
  14. EMBO J. v.10 Mello, C.C.;Kramer, J.M.;Stinchcomb, D.;Ambros, V.
  15. Genet. Eng. Res. v.8 Min, K.M.;Choi, J.D.;Cho, N.J.
  16. Annu. Rev. Neurosci. v.10 Nathanson, N.M. https://doi.org/10.1146/annurev.ne.10.030187.001211
  17. Nature v.334 Peralta, E.G.;Ashkenazi, A.;Winslow, J.W.;Ramachandran, J.;Capon, D.J. https://doi.org/10.1038/334434a0
  18. Science v.267 Segalat, L.;Elkes, D.A.;Kaplan, J.M. https://doi.org/10.1126/science.7886454
  19. J. Biol. Chem. v.263 Shapiro, R.A.;Scherer, N.M.;Habecker, B.A.;Subers, E.M.;Nathanson, N.M.
  20. Proc. Natl. Acad. Sci. USA v.86 Shapiro, R.A.;Wakimoto, B.T.;Subers, E.M.;Nathanson, N.M. https://doi.org/10.1073/pnas.86.22.9039
  21. EMBO J. v.7 Stein, R.;Pinkas-Kramarski, R.;Sokolovsky. M.
  22. J. Biol. Chem. v.265 Tietje, K.M.;Goldman, P.S.;Nathanson, N.M.
  23. Nature v.235 Yatani, A.;Codina, J.;Brown, A.M.;Birnbaumer, L.