Summary
-
1.
The binding of substance P (SP) to synaptic vesicles from rat brain was studied by use of the 125I-Tyr8-analogue of SP.
-
2.
The pH dependence of the binding of both peptides to the lipid extractable fraction of synaptic vesicles was shown to be comparable.
-
3.
The binding of 125I-Tyr8-SP shows a rate constant of association (k 1=6.6×106 M−1 s−1), a rate constant of dissociation (k −1=6.4×10−4 s−1) and gives a K Dof 1×10−10 M. K Dderived from equilibrium studies was 3.2×10−10 M.
-
4.
The binding of 125I-Tyr8-SP to lipids of synaptic vesicles was shown to be reversible, saturable and highly specific.
-
5.
The kinetic data suggest one population of binding sites with a maximal number of 0.8 pmol per mg protein of the synaptic vesicle preparation.
-
6.
Unlabeled SP and the (2–11)-, (3–11)- and (4–11)-analogues of SP inhibit the binding of 125I-Tyr8-SP in a decreasing order in a competitive way when added in excess. Tyr8-SP and eledoisin did not interfere with the binding of 125I-Tyr8-SP whereas uperolein and neurotensin caused a partial inhibition. Physalaemin and d-Ala2-d-Met5-enkephalin enhance the binding of 125I-Tyr8-SP in a cooperative way.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abood, L. G., Hoss, W. P.: Stereospecific binding of morphine to phosphatidylserine. Psychopharmacol. Communications 1, 29–35 (1975)
Chang, M. M., Leeman, S. E.: Isolation of a sialogogic peptide from bovine hypothalamic tissue and its characterisation as substance P. J. Biol. Chem. 245, 4784–4790 (1970)
Cuello, A. C., Jessel, T. M., Kanazawa, I., Iversen, L. L.: Substance P: Localization in synaptic vesicles in rat central nervous system. J. Neurochem. 29, 747–751 (1977)
Cuello, A. C., Emson, P., Del Fiacco, M., Gale, J., Iversen, L. L., Jessel, T. M., Kanazawa, J., Paxinos, G., Quik, M.: Distribution and release of substance P in the central nervous system. In: Centrally acting peptides, (Hughes ed.) London: Macmillan. 1978
Davis, M. E., Akera, T., Brody, T. M., Watson, L.: Opiate receptor: Cooperativity of binding observed in brain slices. Proc. Natl. Acad. Sci. USA 74, 5764–5766 (1977)
DeRobertis, E., Rodriguez de Lores Arneiz, G., Salganicoff, L., Pellegrino de Iraldi, A., Zieher, L. M.: Isolation of synaptic vesicles and structural organisation of the acetylcholine system within brain nerve endings. J. Neurochem. 10, 225–235 (1963)
Devynck, M. A., Meyer, P.: Angiotensin receptors. Biochem. Pharmacol. 27, 1–5 (1978)
Glossmann, H., Baukal, A. J., Catt, K. J.: Properties of angiotensin II receptors in the bovine and rat adrenal cortex. J. biol. Chem. 249, 825–834 (1974)
Heizmann, A., Lembeck, F., Seidel, G.: Die Gewinnung von Substanz P aus dem Lipidextract von Hirngewebe. Naunyn-Schmiedebergs Arch. Exp. Path. Pharmak. 253, 265–279 (1966)
Hökfelt, T., Johansson, O., Kellerth, J. O., Ljungdahl, A., Nilsson, G., Nygards, A., Pernow, B.: Immunohistochemical distribution of substance P. In: Substance P, (von Euler, U.S., Pernow, B., eds.). New York: Raven Press 1977
Inouye, A., Kataoka, K.: Sub-cellular distribution of the substance P in the nervous tissues. Nature (Lond.) 193, 585 (1962)
Jessel, T., Iversen, L. L., Kanazawa, J.: Release and metabolism of substance P in rat hypothalamus. Nature (Lond.) 264, 81–83 (1976)
Johnson, D. A., Cho, T. M., Loh, H. H.: The binding of 5-hydroxytryptamine to acidic lipids in isobutanol. J. Neurochem. 29, 1101–1103 (1977a)
Johnson, D. A., Cho, T. M., Loh, H. H.: Identification of 5-hydroxytryptamine binding substances from rat brain stem. J. Neurochem. 29, 1105–1109 (1977b)
Johnson, D. A., Merlone, S. C., Loh, H. H., Ellman, G. L.: Binding of 5-hydroxytryptamine to acidic lipids in an aqueous medium. J. Neurochem. 31, 713–717 (1978)
Kitabgi, P., Carraway, R., Van Rietschoten, J., Granier, C., Morgat, J. L., Menez, A., Leeman, S., Freychet, P.: Neurotensin: specific binding to synaptic membranes from rat brain. Proc. Natl. Acad. Sci. USA 74, 1846–1850 (1977)
Lembeck, F., Holasek, A.: Die intracelluläre Lokalisation der Substanz P. Naunyn-Schmiedebergs Arch. Exp. Path. Pharmak. 238, 542–545 (1960)
Lembeck, F., Mayer, N., Schindler, G.: Substance P in rat brain synaptosomes. Nauny-Schmiedeberg's Arch. Pharmacol 301, 17–22 (1977)
Lembeck, F., Mayer, N., Schindler, G.: Substance P: binding to lipids in the brain. Naunyn-Schmiedeberg's Arch. Pharmacol. 303, 79–86 (1978)
Loh, H. H., Cho, T. M., Wu, Y., Way, E.: Stereospecific binding of narcotics to brain cerebrosides. Life Sci. 14, 2231–2245 (1974)
Loh, H. H., Law, P. Y., Ostwald, T., Cho, T. H., Way, E. L.: Possible involement of cerebroside sulfate in opiate receptor binding. Fed. Proc. 37, 147–152 (1978)
Lowry, O. H., Rosebrough, N. J., Farr, A. L., Randall, R. J.: Protein measurement with the folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)
Mahler, H. R., Cordes, E. H.: Biological Chemistry. New York: Harper and Row Publishers; San Francisco-London: Evanston, 1971
Miller, R. J., Chang, K. J., Leighton, J., Cuatrecasas, P.: Interaction of iodinated enkephalin analogues with opiate receptors. Life Sci. 22, 379–388 (1978)
Mroz, E. A., Brownstein, M. J., Leeman, S. E.: Distribution of immunoassayable Substance P in the rat brain. Evidence for the existance of substance P-containing tracts. In: Substance P (von Euler, U.S., Pernow, B., eds). New York: Raven Press 1977
Nakata, Y., Kusaka, Y., Segawa, T., Yajima, H., Kitagawa, K.: Substance P: Regional distribution and specific binding to synaptic membranes in rabbit central nervous system. Life Sci. 22, 259–268 (1978)
Otsuka, M., Konishi, S.: Release of substance P-like immunoreactivity from isolated spinal cord of the newborne rat. Nature (Lond.) 264, 83–84 (1976)
Pickel, V. M., Reis, D. J., Leeman, S. E.: Ultrastructural localization of substance P in neurons in rat spinal cord. Brain Res. 122, 534–540 (1977)
Ryall, R. W.: The subcellular distributions of acetylcholine, substance P, 5-hydroxytryptamine, γ-aminobutyric acid and glutamic acid in brain homogenates. J. Neurochem. 11, 131–145 (1964)
Scatchard, G.: The attraction of proteins for small molecules and ions. Ann. N. Y. Acad. Sci. 51, 660 (1949)
Schenker, C., Mroz, E. A., Leeman, S. E.: Release of substance P from isolated nerve endings. Nature (Lond.) 264, 790–792 (1976)
Simantov, R., Childers, S. R., Snyder, S. H.: The opiate receptor binding interactions of 3H-methionin enkephalin, an opioid peptide. Eur. J. Pharmacol. 47, 319–331 (1978)
Uhl, G., Snyder, S. H.: Neurotensin receptor binding, regional and subcellular distribution favor transmitter role. Eur. J. Pharmacol. 41, 89–91 (1977)
Author information
Authors and Affiliations
Additional information
The investigation was supported by grant No. 3506 from the Fonds zur Förderung der wissenschaftlichen Forschung Österreichs and by grant No. 1293 from the Jubiläumsfonds der Österreichischen Nationalbank
Rights and permissions
About this article
Cite this article
Mayer, N., Lembeck, F., Saria, A. et al. Substance P: Characteristics of binding to synaptic vesicles of rat brain. Naunyn-Schmiedeberg's Arch. Pharmacol. 306, 45–51 (1979). https://doi.org/10.1007/BF00515592
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00515592