Lopez-Corcuera B, Kanner B I, Aragón C
Departamento de Biología Molecular, Facultad de Ciencias, Universidad Autónoma, Madrid, Spain.
Biochim Biophys Acta. 1989 Aug 7;983(2):247-52. doi: 10.1016/0005-2736(89)90240-x.
The (Na+ + Cl-)-coupled glycine transporter has been solubilized from rat spinal cord with 2% cholate and purified 6-7-fold using Wheat Germ Agglutinin-Sepharose 4B. Transport activity - as determined upon reconstitution of the fraction into liposomes - was retained on the column and eluted by N-acetylglucosamine. When the glycoprotein fraction was depleted of the N-acetylglucosamine and applied to a second round of lectin-chromatography, the glycine transport activity was retained and again could be eluted by the sugar. The transporter activity reconstituted from the glycoprotein fraction retains the same features displayed in the synaptic plasma membrane vesicles, namely an absolute dependence on sodium and chloride, electrogenicity and efflux and exchange properties. These observations indicate that the (Na+ + Cl-)-coupled glycine transporter is a glycoprotein.
（Na⁺ + Cl⁻）偶联的甘氨酸转运体已用2%胆酸盐从大鼠脊髓中溶解出来，并使用麦胚凝集素 - 琼脂糖4B纯化了6 - 7倍。转运活性（在将该组分重构到脂质体中时测定）保留在柱上，并被N - 乙酰葡糖胺洗脱。当糖蛋白组分中的N - 乙酰葡糖胺被去除并应用于第二轮凝集素色谱时，甘氨酸转运活性得以保留，并且再次可以被该糖洗脱。从糖蛋白组分重构的转运体活性保留了突触质膜小泡中显示的相同特征，即对钠和氯的绝对依赖性、电生性以及外流和交换特性。这些观察结果表明，（Na⁺ + Cl⁻）偶联的甘氨酸转运体是一种糖蛋白。
