Solubilization and assay of [ 3H]imipramine binding sites from human platelets

Abstract

[ 3H]Imipramine binding sites were characterized on outdated (72–96 h) human platelets. The binding site profile was very similar to that previously described for fresh platelets and for CNS membrane preparations. [ 3H]Imipramine binding sites are believed to represent the recognition site for the 5-hydroxytryptamine uptake system. In order to further investigate this molecular relationship, we solubilized [ 3H]imipramine binding sites using 1% digitonin and characterised them using a polyethylene glycol precipitation assay. Solubilization did not alter the pharmacological specificity or affinity of the [ 3H]imipramine binding site and recovery of sites was very high. It was concluded that the solubilization procedure left the binding site remarkably intact. The ready availability of platelets and the convenience of the binding assay makes this system amenable for further purification steps.