Partial purification of [3H]mianserin binding sites.

Frontal cortex membranes were solubilized with digitonin, prelabelled with 4 nM [3H]mianserin, and partially purified by isoelectric focussing. Bound [3H]mianserin separated from free [3H]mianserin as a single radioactive peak with a pI value of 5.03. A 14-fold purification was achieved. Focussing in the presence of 1 microM ketanserin (R 41468) or 1 microM chlorpyramine (a histamine H1-antagonist) or 10 microM spiperone (an S2-antagonist) completely abolished the peak of radioactivity at pH 5. Pharmacological characterization of the eluted radioactive peak revealed both histaminergic and serotonergic binding properties. The data suggest that the serotonergic and histaminergic components of [3H]mianserin binding to frontal cortex are not separable by solubilization, gel filtration or isoelectric focussing.