Solubilization of rat brain serotonin-S2 receptors using CHAPS/salt.
Serotonin-S2 receptors from rat frontal cortex were solubilized with a mixture of 6.8 mM CHAPS (3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulphonate) and 1.4 M sodium chloride. This new solubilization procedure solubilized about 40% of the membrane-bound serotonin-S2 receptors in an active form. The solubilized receptors were not sedimented after 1 h of centrifugation at 100 000 X g and they passed freely through 0.20 micron filters. The solubilized preparation showed high affinity binding of [3H]ketanserin and revealed a typical serotonin-S2 receptor profile: binding could be displaced by nanomolar concentrations of different serotonin antagonists and by micromolar concentrations of serotonin agonists. Compounds belonging to other pharmacological classes were poorly, or not active. Upon density gradient sedimentation, Svedberg coefficients of approximately 5 S were found on sucrose gradients made with H2O or D2O as the solvent. This was much lower than the value of 11.5 S previously reported from lysophosphatidylcholine-solubilized serotonin-S2 receptors.
Wouters
W
W
Van Dun
J
J
Leysen
J E
JE
Laduron
P M
PM
D000818:Animals; D001923:Brain Chemistry; D002499:Centrifugation, Density Gradient; D002793:Cholic Acids; D005374:Filtration; D066298:In Vitro Techniques; D007650:Ketanserin; D008861:Microsomes / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000302:isolation & purification; D010880:Piperidines / Q000378:metabolism; D051381:Rats; D011919:Rats, Inbred Strains; D011985:Receptors, Serotonin / Q000302:isolation & purification* / Q000378:metabolism; D012702:Serotonin Antagonists / Q000494:pharmacology; D012965:Sodium Chloride
D002793:Cholic Acids; D009419:Nerve Tissue Proteins; D010880:Piperidines; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; D012965:Sodium Chloride; D007650:Ketanserin; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate