Cash C D, Hechler V, Mersel M, Maitre M
L.N.M.I.C UPR-416 CNRS, Centre de Neurochimie, Strasbourg, France.
Neurosci Lett. 1996 May 3;209(1):25-8. doi: 10.1016/0304-3940(96)12589-1.
The solubilisation of the gamma-hydroxybutyrate (GHB) receptors from rat brain membranes was undertaken as the first step for their molecular characterisation and purification. Treatment of crude brain membranes with high concentrations of NaCl and Triton X-100 resulted in solubilisation of proteins which retain specific GHB binding activity. Ionic detergents do not solubilise and/or inactivate the receptors. Measurements of kinetic parameters of GHB binding showed that the solubilised receptor, in the presence of detergent, exhibited a reduction of affinity for GHB and its endogenous brain analogue trans-4-hydroxycrotonate (T-HCA). The membrane protein extract, submitted to chromatography by gel filtration, showed a single peak of protein with [3H]GHB binding activity. Association and dissociation constants of GHB for its membrane binding site were in accordance with the Kd determined by the Scatchard method.
从大鼠脑膜中溶解γ-羟基丁酸酯（GHB）受体是对其进行分子表征和纯化的第一步。用高浓度的氯化钠和 Triton X-100 处理粗脑膜会导致保留特定 GHB 结合活性的蛋白质溶解。离子去污剂不会溶解和/或使受体失活。对 GHB 结合动力学参数的测量表明，在去污剂存在的情况下，溶解的受体对 GHB 及其内源性脑类似物反式-4-羟基巴豆酸酯（T-HCA）的亲和力降低。经凝胶过滤色谱分析的膜蛋白提取物显示出一个具有[3H]GHB 结合活性的单一蛋白峰。GHB 与其膜结合位点的缔合常数和解离常数与通过 Scatchard 方法测定的 Kd 一致。
