Wouters W, van Dun J, Laduron P M
Biochem Pharmacol. 1984 Dec 15;33(24):4039-44. doi: 10.1016/0006-2952(84)90018-2.
[3H]Spiperone binding sites were solubilized in high yield from human, dog and rat brain with a mixture of sodium cholate (0.3% w/v) and sodium chloride (1.4 M). The binding sites were not sedimented after one hour at 100,000 g, they passed freely through 0.20 micron filters, migrated as a single peak in gradient sedimentation and were retarded upon gel filtration, proving that they were truly solubilized. The solubilized binding sites were definitely of dopaminergic nature. They showed saturable, reversible, high affinity binding of [3H]spiperone; displacement of [3H]spiperone binding by nanomolar concentrations of dopamine antagonists and micromolar concentrations of serotonin antagonists; stereo-specificity and a good correlation with drug affinities for membrane preparations. The non-displaceable, non-specific [3H]spiperone binding was very low. Gradient sedimentation analysis revealed a sedimentation coefficient of 12 S for dog solubilized preparations, 9 S for rat solubilized preparations and only 2.5 S for human solubilized preparations (values, uncorrected for detergent binding). Gel filtration experiments seem to confirm these molecular characteristics. Therefore the present results show that the dopamine receptor reveals the same pharmacological properties when solubilized with cholate-salt from rat, dog or human brain, while physico-chemical properties seem to indicate some differences.
用胆酸钠（0.3%，w/v）和氯化钠（1.4M）的混合物可从人、狗和大鼠脑中高效溶解[3H]螺哌隆结合位点。这些结合位点在100,000g离心1小时后不会沉淀，能自由通过0.20微米滤膜，在梯度沉降中呈单一峰迁移，且在凝胶过滤时受阻，证明它们确实已被溶解。溶解的结合位点肯定具有多巴胺能性质。它们表现出对[3H]螺哌隆的可饱和、可逆、高亲和力结合；纳摩尔浓度的多巴胺拮抗剂和微摩尔浓度的5-羟色胺拮抗剂能取代[3H]螺哌隆的结合；具有立体特异性，且与药物对膜制剂的亲和力有良好相关性。不可取代的非特异性[3H]螺哌隆结合非常低。梯度沉降分析显示，狗溶解制剂的沉降系数为12S，大鼠溶解制剂为9S，人溶解制剂仅为2.5S（该值未校正去污剂结合）。凝胶过滤实验似乎证实了这些分子特征。因此，目前的结果表明，当用胆酸盐从大鼠、狗或人脑中溶解时，多巴胺受体显示出相同的药理学特性，而物理化学性质似乎表明存在一些差异。
