Solubilization and characterization of the 5-hydroxytryptamine transporter complex from rat cerebral cortical membranes.
The 5-hydroxytryptamine transporter complex from rat cerebral cortical membranes was solubilized with digitonin. The affinity of the solubilized transporter complex for [3H]paroxetine, a very selective and potent inhibitor of 5-hydroxytryptamine uptake, was not affected and remained unchanged when compared with the parent membrane preparation. The solubilization yield of membrane-bound [3H]paroxetine binding sites was 42%. The pharmacological profile of the solubilized transporter complex was similar to that of the intact transporter in membranes of the cerebral cortex, with the exception of tryptamine, which exhibited a 10-fold loss in potency to inhibit [3H]paroxetine binding to the solubilized transporter when compared to membranes. The Stokes radius determined by gel filtration was 7.6 nm. This successful solubilization of the neuronal 5-hydroxytryptamine transporter complex is the starting point for purification of this macromolecular moiety.
D000818:Animals; D001692:Biological Transport; D002540:Cerebral Cortex / Q000032:analysis* / Q000378:metabolism; D002850:Chromatography, Gel; D007700:Kinetics; D008297:Male; D008566:Membranes / Q000032:analysis; D017374:Paroxetine; D010880:Piperidines / Q000494:pharmacology; D051381:Rats; D012701:Serotonin / Q000032:analysis* / Q000378:metabolism
D010880:Piperidines; D012701:Serotonin; D017374:Paroxetine