The binding of acidic amino acids to snail, Helix aspersa, periesophagic ring membranes reveals a single high-affinity glutamate/kainate site.
The characterization of specific acidic amino acid binding sites to snail, Helix aspersa, ganglia membranes has been assayed using tritiated glutamate (L-[3H]Glu), aspartate (L-[3H]Asp), cysteine sulfinate (L-[3H]CSA) and kainate. At 2 degrees C, only L-[3H]Glu and [3H]kainate specific binding could be measured using a filtration procedure to separate bound from free ligand. The analysis of L-[3H]Glu specific binding reveals the presence of one class of high-affinity binding sites with Kd = 0.12 microM and Bmax = 30 pmol/mg protein. This L-[3H]Glu binding was specific, reversible and saturable. The order of potency of different substances, agonists or antagonists of the rat brain excitatory amino acid receptors, has been determined. Kainate was the best displacing agent, followed by ibotenate = L-Glu greater than L-alpha-aminoadipate (L-alpha-AA) greater than homocysteate (HCA). Using 10 nM [3H]kainate, a single class of binding site was detected. Its pharmacological properties indicate that it is likely identical to the L-[3H]Glu binding site. This L-Glu-kainate site possesses most of the properties expected for a specific receptor. However, whereas L-[3H]Glu binding could be detected on purified neuronal membranes, the major component of specifically bound L-[3H]Glu appeared to be located on the sheaths surrounding neuronal cell bodies. These findings suggest that Glu or another endogenous acidic amino acid may function as a transmitter at neuromuscular junctions in Helix periesophagic ring, acting at a receptor distinct from those on nerve cells.
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Bockaert
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Recasens
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D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D001665:Binding Sites; D003545:Cysteine / Q000031:analogs & derivatives / Q000378:metabolism; D005724:Ganglia / Q000378:metabolism*; D005971:Glutamates / Q000378:metabolism; D018698:Glutamic Acid; D006372:Helix, Snails; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D007608:Kainic Acid / Q000378:metabolism; D007700:Kinetics; D018377:Neurotransmitter Agents; D009994:Osmolar Concentration
D000596:Amino Acids; D005971:Glutamates; D018377:Neurotransmitter Agents; D001224:Aspartic Acid; D018698:Glutamic Acid; D003545:Cysteine; D007608:Kainic Acid; C013461:cysteine sulfinic acid