title: Binding of N-[propionyl-3H]propionylated alpha-bungarotoxin and L-[benzilic-4,4'-3H] quinuclidinyl benzilate to CNS extracts of the cockroach Periplaneta americana.
abstract: The nerve cord of the cockroach (Periplaneta americana) contains distinct saturable components of specific binding for the ligands N-[propionyl-3H]propionylated alpha-bungarotoxin and L-[benzilic-4,4'-3H]quinuclidinyl benzilate. N-[Propionyl-3H]propionylated alpha-bungarotoxin bound reversibly to homogenates with a Kd of 4.8 nM and Bmax of 910 fmol mg-1. The association rate constant (1.9 X 10(5) M-1 s-1) and dissociation rate constant (1.2 X 10(-4) s-1) yielded a Kd of 0.6 nM. Nicotinic ligands were found to displace toxin binding most effectively. The binding sites characterized in this way showed many similarities with the properties of the vertebrate neuronal alpha-bungarotoxin binding site. For a range of cholinergic ligands, inhibition constants calculated from toxin binding studies closely corresponded to their effectiveness in blocking the depolarizing response to acetylcholine recorded by electrophysiological methods from an identified cockroach motoneurone. The N-[propionyl-3H]propionylated alpha-bungarotoxin binding component therefore appears to be a constituent of a functional CNS acetylcholine receptor. Binding of L-[benzilic-4,4'-3H]quinuclidinyl benzilate was reversible with a Kd of 8 nM and Bmax of 138 fmol mg-1, determined from equilibrium binding experiments. The Kd calculated from the association rate constant (2.4 X 10(5) M-1 s-1) and dissociation rate constant (1.3 X 10(-4) s-1) was 1.9 nM. Muscarinic ligands were the most potent inhibitors of quinuclidinyl benzilate binding. The characteristics of this binding site resembled those of vertebrate CNS muscarinic cholinergic receptors. In contrast with vertebrate CNS, the nerve cord of Periplaneta americana contains more (approximately X 7) alpha-bungarotoxin binding sites than quinuclidinyl benzilate binding sites.
authors.forename: S C
authors.initials: SC
authors.lastname: Lummis
authors.forename: D B
authors.initials: DB
authors.lastname: Sattelle
mesh_terms: D000818:Animals; D001667:Binding, Competitive / Q000187:drug effects; D002038:Bungarotoxins / Q000378:metabolism*; D002490:Central Nervous System / Q000378:metabolism; D002645:Chickens; D003058:Cockroaches / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000378:metabolism; D006110:Grasshoppers / Q000378:metabolism; D006168:Guinea Pigs; D006624:Hippocampus / Q000378:metabolism; D007700:Kinetics; D008297:Male; D051379:Mice; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009447:Neuroblastoma / Q000378:metabolism; D010526:Periplaneta / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D011812:Quinuclidines / Q000378:metabolism*; D011813:Quinuclidinyl Benzilate / Q000031:analogs & derivatives / Q000378:metabolism*; D011950:Receptors, Cholinergic / Q000378:metabolism*
chemical_list: C044493:1-benzilic-4,4'-quinuclidinyl benzilate; D002038:Bungarotoxins; D009419:Nerve Tissue Proteins; D011812:Quinuclidines; D011950:Receptors, Cholinergic; C030937:propionyl-alpha-bungarotoxin; D011813:Quinuclidinyl Benzilate