Characteristics of 3H-substance P binding sites in rat brain membranes.
Binding characteristics of 3H-Substance P (SP) were studied with rat brain membranes using a method applied to peripheral tissues by Lee and Snyder [15]. This method was well applicable to central nervous system (CNS) tissues. The results in the present study indicate that specific 3H-SP binding reaches a plateau only after 20 minutes of incubation, and the binding sites are saturable at a relatively low concentration of 3H-SP. Scatchard analysis of specific binding data reveals a single class of binding sites with a high affinity (Kd = 0.30 nM) and a low density (Bmax = 27.7 fmol/mg protein) in rat brain membranes. A Hill plot of the displacement curve of 3H-SP with unlabelled SP showed no indication for cooperativity (nH = 0.83). The relative potencies of binding of various SP fragments at 3H-SP binding sites were fairly parallel to the length of the C-terminal fragments. Neurotransmitters not structurally related to SP produced no effect on 3H-SP binding even when used at micromolar concentrations.
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D007700:Kinetics; D008297:Male; D051381:Rats; D011919:Rats, Inbred Strains; D018040:Receptors, Neurokinin-1; D017981:Receptors, Neurotransmitter / Q000378:metabolism*; D013373:Substance P / Q000378:metabolism*; D014316:Tritium
D018040:Receptors, Neurokinin-1; D017981:Receptors, Neurotransmitter; D014316:Tritium; D013373:Substance P