Dicyclohexylcarbodiimide inhibits the monoamine carrier of bovine chromaffin granule membrane.
The monoamine carrier of bovine chromaffin granule membrane catalyzes a H+/neutral amine antiport. Dicyclohexylcarbodiimide (DCCD) inhibits this carrier in a time- and concentration -dependent manner as shown by the following evidence: it inhibits the carrier-mediated pH gradient driven monoamine uptake without collapsing the pH gradient; it affects the binding of the specific inhibitors [2-3H]dihydrotetrabenazine and [3H]reserpine. The DCCD inhibition of the carrier occurs in the same concentration range as that of the ATP-dependent H+ translocase. Saturation isotherms of [2-3H]dihydrotetrabenazine binding indicate that DCCD decreases the number of binding sites without any change of the equilibrium dissociation constant. Kinetic studies of DCCD inactivation indicate that the modification of only one amino acid residue is responsible for the inhibition. Preincubation of the membranes with tetrabenazine protects the carrier against inactivation by DCCD: in this case, [2-3H] dihydrotetrabenazine binding and pH gradient driven monoamine uptake are restored after washing out of DCCD and tetrabenazine. We suggest the existence in the monoamine carrier of a carboxylic acid involved in H+ translocation, similar to those demonstrated not only in F0-F1 ATPases but also in cytochrome c oxidase, mitochondrial cytochrome b-c1 complex, and nucleotide transhydrogenase. Protonation-deprotonation of this group would affect the binding of [2-3H]dihydrotetrabenazine by the carrier.
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000588:Amines / Q000378:metabolism*; D001667:Binding, Competitive; D002234:Carbodiimides / Q000494:pharmacology*; D002837:Chromaffin Granules / Q000378:metabolism*; D002838:Chromaffin System / Q000378:metabolism*; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology*; D006863:Hydrogen-Ion Concentration; D007425:Intracellular Membranes / Q000187:drug effects / Q000378:metabolism*; D007700:Kinetics; D012110:Reserpine / Q000378:metabolism; D012701:Serotonin / Q000378:metabolism; D013747:Tetrabenazine / Q000031:analogs & derivatives / Q000378:metabolism; D014439:Tyramine / Q000378:metabolism
D000588:Amines; D002234:Carbodiimides; D012701:Serotonin; C032811:dihydrotetrabenazine; D004024:Dicyclohexylcarbodiimide; D012110:Reserpine; D000251:Adenosine Triphosphatases; D014439:Tyramine; D013747:Tetrabenazine