Partial purification and characterization of the sodium-ion-coupled 5-hydroxytryptamine transporter of rat cerebral cortex.
A procedure for the extensive purification of the Na(+)-coupled 5-hydroxytryptamine transporter of rat cerebral cortex has been developed. The 5-hydroxytryptamine transporter was solubilized with the non-ionic detergent digitonin, and the detergent extracts were subjected to sequential affinity chromatography on a citalopram-based agarose support and wheat-germ-agglutinin-Sepharose. 5-Hydroxytryptamine transporters in the affinity-purified preparation were identified by using the selective 5-hydroxytryptamine-uptake inhibitor [3H]paroxetine, and were shown to display a similar pharmacological profile to those present in particulate preparations. An overall transporter purification of around 2000-fold was achieved with a 9% recovery. SDS/PAGE of affinity-chromatographed material starting from detergent extracts incubated in the presence or absence of 1 mM-citalopram indicated that a polypeptide of M(r) 73,000 corresponded to the 5-hydroxytryptamine-transporter protein.
Graham
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Synthélabo Recherche (L.E.R.S.), Bagneux, France.
Esnaud
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H
Langer
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D000818:Animals; D002352:Carrier Proteins / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism; D002414:Cations, Monovalent; D002540:Cerebral Cortex / Q000737:chemistry* / Q000378:metabolism; D002846:Chromatography, Affinity; D003902:Detergents; D004072:Digitonin; D004591:Electrophoresis, Polyacrylamide Gel; D008562:Membrane Glycoproteins / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D017374:Paroxetine / Q000494:pharmacology; D051381:Rats; D012701:Serotonin / Q000378:metabolism*; D012702:Serotonin Antagonists / Q000494:pharmacology; D050486:Serotonin Plasma Membrane Transport Proteins; D012964:Sodium / Q000378:metabolism*
D002352:Carrier Proteins; D002414:Cations, Monovalent; D003902:Detergents; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D012702:Serotonin Antagonists; D050486:Serotonin Plasma Membrane Transport Proteins; C493232:Slc6a4 protein, rat; D012701:Serotonin; D017374:Paroxetine; D012964:Sodium; D004072:Digitonin