[3H]5-hydroxytryptamine binding to reconstituted fraction with sulphatides, phosphatidylserine and phosphatidylinositol.
As a first step toward the examination of the involvement of sulphatides, phosphatidylserine and phosphatidylinositol in 5-HT receptor mechanisms, we performed [3H]5-HT binding experiments on the various reconstituted fractions with these acidic lipids. A binding assay of [3H]5-HT to these fractions was carried out by Sephadex LH20 column chromatography. Among various reconstituted fractions, only the reconstitution system with the three acidic lipids exhibited a saturable [3H]5-HT binding capacity, whereas no binding was seen with [3H]-spiperone. When the binding of [3H]5-HT to this fraction was plotted as a function of the ligand concentration, a multiple binding mode with three classes of binding components (or sites) was observed. Furthermore, the double reciprocal plot indicated that this reconstitution system had three apparent KD values of 4.7, 15 and 59 nM. The displacement studies with various compounds indicated that only a few 5-HT agonists (5-methoxytryptamine and tryptamine) and neurotransmitters (DA and ACh) inhibited the [3H]5-HT binding to this fraction, but 5-HT antagonists, LSD analogues and neuroleptics had no effect. Moreover, GTP, GDP and Gpp(NH)p clearly inhibited the [3H]5-HT binding in spite of their weak potencies, while GMP did not have any effect.
D000818:Animals; D001921:Brain / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D066298:In Vitro Techniques; D008297:Male; D008563:Membrane Lipids / Q000378:metabolism; D010716:Phosphatidylinositols / Q000378:metabolism*; D010718:Phosphatidylserines / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D012701:Serotonin / Q000378:metabolism*; D013433:Sulfoglycosphingolipids / Q000378:metabolism*; D013570:Synaptic Membranes / Q000378:metabolism
D008563:Membrane Lipids; D010716:Phosphatidylinositols; D010718:Phosphatidylserines; D013433:Sulfoglycosphingolipids; D012701:Serotonin