Summary
-
1.
The partition of substance P (SP) between buffer solutions (pH 1.6–7.8) and an organic, phospholipid (phosphatidyl serine, phosphatidyl ethanolamine, phosphatidyl inositol and phosphatidyl choline) containing phase (chloroform:methanol 2:1) was studied.
-
2.
The binding of SP to phosphatidyl serine, phosphatidyl ethanolamine and phosphatidyl inositol was lowest at pH 2 and increased with pH. The binding to phosphatidyl choline was much smaller and less dependent on pH.
-
3.
In contrast to the basic peptide SP (pI 10.5), physalaemin (pI 7.0) did not show any binding to phospholipids at any investigated pH value which underlines the importance of a basic group in the peptide for its binding.
-
4.
The high affinity (K D =0.1 μM) and capacity of 44 pmol SP/μg phosphatidyl serine and 48 pmol SP/μg phosphatidyl ethanolamine at pH 7.2 under conditions of saturation contrasted with the very low binding of SP to phosphatidyl inositol or phosphatidyl choline. Ionic bindings between the basic peptide and phosphatidyl serine or phosphatidyl ethanolamine are regarded to be predominant, although other binding forces cannot be excluded.
-
5.
There was a concentration-dependent reduction in the binding of SP to phosphatidyl serine or phosphatidyl ethanolamine by Na+ and Ca2+, whereas K+ showed hardly any effect at physiological concentrations.
-
6.
The model studies served to consider the possibilities of the binding of a basic peptide to lipid storage or receptor sites.
Similar content being viewed by others
References
Abood, L. G., Hoss, W. P.: Stereospecific binding of morphine to phosphatidyl serine. Psychopharmacol. Comm. 1, 29–35 (1975)
Abood, L. G., Takeda, F.: Enhancement of stereospecific opiate binding to neural membranes by phosphatidyl serine. Eur. J. Pharmacol. 39, 71–77 (1976)
Cuello, A. C., Jessell, T. M., Kanazawa, J., Iversen, L. L.: Substance P: localization in synaptic vesicles in rat central nervous system. J. Neurochem. 29, 747–781 (1977)
Eber, O., Lembeck, F.: Über den enzymatischen Abbau der Substanz P. Naunyn-Schmiedebergs Arch. exp. Path. Pharmakol. 229, 139–147 (1956)
Geipert, F., Lembeck, F., Sprössler, B.: Differentiation of substance P and physalaemin. Nauny-Schmiedebergs Arch. exp. Path. Pharmakol. 265, 225–232 (1969)
Hauser, H., Darke, A., Phillips, M. C.: Ion-binding to phospholipids. Interaction of calcium with phosphatidylserine. Eur. J. Biochem. 62, 335–344 (1976)
Hauser, H., Finer, E. G., Darke, A.: Crystalline anhydrous Caphosphatidylserin bilayers. Biochem. Biophys. Res. Commun. 76, 267–274 (1977)
Heizmann, A., Lembeck, F., Seidel, G.: Die Gewinnung der Substanz P aus dem Lipidextrakt von Hirngewebe. Naunyn-Schmiedebergs Arch. exp. Path. Pharmakol. 253, 265–279 (1966)
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)
Lee, C. Y., Akera, T., Brody, T. M.: Effects of Na+, K+, Mg2+ and Ca2+ on the saturable binding of 3H-dihydromorphine and 3H-naloxone in vitro. J. Pharmacol. Exp. Ther. 202, 166–173 (1977)
Lembeck, F.: Substance P: Binding to lipids in brain. In: Symposium on Biologically Acting Peptides (J. Hughes and H. W. Kosterlitz, eds.), pp. 124–134. New York: Macmillan Press 1978
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, D. Y., Ostwald, T., Cho, T. M., Way, E. L.: Possible involvement of cerebroside sulfate in opiate receptor binding. Fed. Proc. 37, 147–152 (1978)
Nagy, A., Baker, R. R., Morris, S. J., Whittaker, V. P.: The preparation and characterization of synaptic vesicles of high purity. Brain Res. 109, 285–309 (1976)
Pert, O., Snyder, S.: Opiate receptor binding of agonists and antagonists affected differentially by sodium. Mol. Pharmacol. 10, 868–870 (1974)
Pickel, V. M., Reis, D. J., Leeman, S. E.: Ultrastructural localization of substance P in neurons of rat spinal cord. Brain Res. 122, 534–540 (1977)
Simon, E. J., Hiller, J. M.: In vitro studies on opiate receptors and their ligands. Fed. Proc. 37, 141–146 (1978)
Simon, E. J., Hiller, J. M., Edelman, J.: Stereospecific binding of the potent narcotic analgesic 3H-etorphine to rat brain homogenate. Proc. Natl. Acad. Sci. USA 70, 1947–1950 (1973)
Sundaralingam, M.: Molecular structures and conformations of the phospholipids and sphingomyelins. Ann. N.Y. Acad. Sci. 195, 324–355 (1972)
Author information
Authors and Affiliations
Additional information
This study was supported by the Fonds zur Förderung der wissenschaftlichen Forschung Österreichs (project No. 3400) and by the Jubiläumsfonds der österreichischen Nationalbank (project No. 1293)
Rights and permissions
About this article
Cite this article
Lembeck, F., Saria, A. & Mayer, N. Substance P: Model studies of its binding to phospholipids. Naunyn-Schmiedeberg's Arch. Pharmacol. 306, 189–194 (1979). https://doi.org/10.1007/BF00507102
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00507102