Structure-Function Relationships of Endothelins, Sarafotoxins, and Their Receptor Subtypes


  • Mordechai Sokolovsky

    Corresponding author
    1. Laboratory of Neurochemistry, Department of Biochemistry, The George S. Wise Faculty of Life Science, Tel Aviv University, Tel Aviv, Israel
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Address correspondence and reprint requests to Dr. M. Sokolovsky at Department of Biochemistry. Tel Aviv University. Tel Aviv 69978. Israel.


Abstract: In brain synaptic membranes not extensively washed, (+)-5-[3H]methyl-10,1 l-dihydro-5H-dibenzo[a, d]-cyclohepten-5,10-imine ([3H]MK-801) binding was markedly inhibited in a concentration-dependent manner (at concentrations above 1 μM) by several compounds having antagonistic activity at the Ca2+-binding protein calmodulin. Scatchard analysis revealed that N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) inhibited the binding through a significant decrease in the density of binding sites without affecting the affinity at 10 μM. In membranes extensively washed and treated with a low concentration of Triton X-100, L-glutamic acid (Glu) drastically accelerated the initial association rate of [3H]MK-801 binding with glycine (Gly), almost doubling the initial association rate found in the presence of Glu alone. The addition of W-7 invariably reduced the initial association rate observed in the presence of either Glu alone or both Glu and Gly, without significantly altering the dissociation rate of bound [3H]-MK-801, irrespective of the presence of the two stimulatory amino acids. The maximal potencies of Glu, Gly, and spermidine in potentiating the binding were all attenuated by W-7. These results suggest that calmodulin antagonists may interfere with opening processes of an ion channel associated with an N-methyl-D-aspartate-sensitive subclass of excitatory amino acid receptors in rat brain.