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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Action of polychlorocycloalkane insecticides on binding of [35S]t-butylbicyclophosphorothionate to Torpedo electric organ membranes and stereospecificity of the binding site.

Binding of [35S]t-butylbicyclophosphorothionate ([35S]TBPS) to Torpedo electric organ membranes was characterized. A dose- and pH-dependent binding (100.8 pmol/mg protein) was detected with a single affinity (Kd of 0.9 microM) in the presence of 150 mM KCl at pH 6. 8. Other anions such as Br- and I- also increased binding affinity, but to a lower degree than Cl-, which increased the affinity by two- to threefold. In presence of 150 mM KCl, [35S]TBPS binding was inhibited noncompetitively by Zn2+ and by 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS) (IC50 of 9 microM). The gamma-isomer of hexachlorocyclohexane ( BHC) was much more potent in inhibiting this [35S]TBPS binding and the inhibition was competitive (Ki = 40 nM). Like binding of [35S]TBPS to the gamma-aminobutyric acid (GABA) receptor, its binding to Torpedo membranes was inhibited by pentobarbital, mephobarbital, and hexobarbital (IC50 of 85, 225, and 300 microM), respectively), but not by phenobarbital. Binding was not inhibited by diazepam, GABA, bicuculline, or avermectin B1a, ligands that bind to the GABAA receptor. [35S]TBPS binding was inhibited by BHC isomers with the following decreasing order of potency alpha = gamma greater than sigma greater than beta, and by cyclodiene insecticides. Endrin was more potent than dieldrin, but endosulfan I and II had similar effects. The data suggest that the binding site for polychlorocycloalkane insecticides on this protein is much less stereoselective than that of the Cl- channel of the GABAA receptor. Also, even though this Torpedo protein has higher affinity for insecticides, such as gamma-BHC, than does the GABAA receptor, it is the latter whose specificity correlates best with polychlorocycloalkane toxicity. Nevertheless, because of its high affinity for gamma-BHC such a protein in muscles or brain may be an important target for the action of this insecticide.[1]


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