Solubilization and characterization of [3H]Imipramine and [3H]Paroxetine binding sites from calf striatum |
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Authors: | A. Rotondo G. Giannaccini C. Quattrone D. Marazziti C. Martin G. B. Cassano A. Lucacchini |
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Affiliation: | (1) Institute of Psychiatry, University of Pisa, via Roma 67, 56100 Pisa, Italy;(2) Instituto Policattedra di Discipline Biologiche, University of Piva, via Roma 67, 56100 Pisa, Italy |
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Abstract: | The serotonin (5-HT) transporter from calf striatum cerebral membranes was solubilized with digitonin and characterized by gel exclusion chromatography. [3H]Imipramine and [3H]paroxetine were utilized as markers for labeling it.3H-imipramine labels a high- and a low-affinity site on striaturn membranes, whereas it binds to a single high-affinity site on the solubilized fraction. [3H]Paroxetine binds with the same affinity to a single site on both membranes and solubilized preparations. After gel exclusion chromatography of the solubilizate both [3H]imipramine and [3H]paroxetine bind on an identical fraction of 205 kDa molecular weight, with a similar maximum number of binding sites (Bmax). Our results suggest that both3H-imipramine and [3H]paroxetine bind to a common site on the 5-HT transporter. |
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Keywords: | Serotonin neuronal serotonin transporter calf striatum [3H]imipramine binding [3H]paroxetine binding purification |
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