Further characterization of the binding of substance P to a fraction from rabbit brain enriched in synaptic membranes
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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
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