Further characterization of the binding of substance P to a fraction from rabbit brain enriched in synaptic membranes
- 12 Downloads
The present experiments were designed to further characterize the specific [3H]substance P (SP) binding to membrane fractions from rabbit brain.
The specific binding was resistant to treatment with proteolytic enzymes whereas phospholipase A (0.7 unit/ml), C (0.025 unit/ml) and D (6.2 unit/ml) reduced the specific binding without decreasing total binding. Neuraminidase (0.01 unit/ml) decreased total binding as well as the specific binding.
High concentration of Triton X-100 reduced the specific binding whereas deoxycholate, at 0.05%, reduced both total and the specific binding.
Na+ (50–200 mM) reduced the specific binding, depending on the concentrations employed, whereas K+ decreased the specific binding at 10 and 100 mM. The binding was reduced considerably by 100 mmol/l of CaCl2.
From these results it is suggested that the specific [3H]SP binding sites in membrane fractions from rabbit brain could be phospholipids.
Key wordsSubstance P Binding sites Phospholipids
Unable to display preview. Download preview PDF.
- De Robertis E, Azcurra JM, Fiszer S (1967) Ultrastructure and cholinergic binding capacity for junctional complexes isolated from rat brain. Brain Res 5:45–56Google Scholar
- Enna SJ, Snyder SH (1977) Influences of ions, enzymes, and detergents on γ-aminobutyric acid-receptor binding in synaptic membranes of rat brain. Mol Pharmacol 13:442–453Google Scholar
- Fiszer S, De Robertis E (1967) Action of Triton X-100 on ultrastructure and membrane-bound enzymes of isolated nerve endings from rat brain. Brain Res 5:31–44Google Scholar
- Lembeck F (1978) Substance P: Binding to lipids in brain. In: Hughes J, Kosterlitz HW (eds) Symposium on biologically acting peptides. Macmillan Press, New York, pp 124–134Google Scholar
- Lembeck F, Mayer N, Schindler G (1978) Substance P: Binding to lipids in the brain. Naunyn-Schmiedeberg's Arch Pharmacol 303:79–86Google Scholar
- Lembeck F, Saria A, Mayer N (1979) Substance P: Model studies of its binding to phospholipids. Naunyn-Schmiedeberg's Arch Pharmacol 306:189–194Google Scholar
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275Google Scholar
- Nakata Y, Kusaka Y, Segawa T, Yajima H, Kitagawa K (1978) Substance P: Regional distribution and specific binding to synaptic membranes in rabbit central nervous system. Life Sci 22:259–268Google Scholar
- Pasternak GW, Snyder SH (1974) Opiate receptor binding: Effects of enzymatic treatments. Mol Pharmacol 10:183–193Google Scholar
- Pasternak GW, Snyder SH (1975) Opiate receptor binding: Enzymatic treatments that discriminate between agonist and antagonist interactions. Mol Pharmacol 11:478–484Google Scholar
- Saria A, Mayer N, Lembeck F, Pabst M (1980) Regional distribution and biochemical properties of 125I-Tyr8-substance P binding sites in synaptic vesicles. Naunyn-Schmiedeberg's Arch Pharmacol 311:151–157Google Scholar
- Segawa T, Nakata Y, Nakamura K, Yajima H, Kitagawa K (1976) Substance P in the central nervous system of rabbits: Uptake system differs from putative transmitters. Jpn J Pharmacol 26:757–760Google Scholar