Gurevich V S, Popov Iu G, Gorodinskiĭ A I, Dambinova S A
Biokhimiia. 1991 Sep;56(9):1632-9.
Solubilization of the total membrane fraction of human platelets in a 2% solution of sodium deoxycholate and subsequent affinity chromatography on glutamate agarose resulted in two protein fractions possessing a glutamate-binding activity. As can be evidenced from radioligand binding data, the first fraction contains two types of binding sites (Kd1 = 1 microM, Bmax 1 = 100 pmol/mg of protein; Kd2 = 9.3 microMm Bmax2 = 395 pmol/mg of protein). The second fraction has only one type of binding sites (Kd = 1 microM, Bmax = = 110 pmol/mg of protein). SDS-PAAG electrophoresis revealed the presence in the first fraction of proteins with Mr of 14, 24, 56 and 155 kDa, whereas the second fraction was found to contain 14, 46, 71 and 155 kDa proteins. Solid phase immunoenzymatic analysis using poly- and monoclonal specific antibodies against mammalian brain glutamate-binding proteins revealed a marked immunochemical similarity of the isolated protein fractions with human brain synaptic membrane glutamate-binding proteins.
将人血小板的总膜组分溶解于2%的脱氧胆酸钠溶液中，随后在谷氨酸琼脂糖上进行亲和层析，得到了两个具有谷氨酸结合活性的蛋白质组分。从放射性配体结合数据可以看出，第一个组分包含两种类型的结合位点（Kd1 = 1微摩尔，Bmax 1 = 100皮摩尔/毫克蛋白质；Kd2 = 9.3微摩尔，Bmax2 = 395皮摩尔/毫克蛋白质）。第二个组分只有一种类型的结合位点（Kd = 1微摩尔，Bmax = 110皮摩尔/毫克蛋白质）。SDS-PAAG电泳显示第一个组分中存在分子量为14、24、56和155 kDa的蛋白质，而第二个组分含有14、46、71和155 kDa的蛋白质。使用针对哺乳动物脑谷氨酸结合蛋白的多克隆和单克隆特异性抗体进行的固相免疫酶分析表明，分离出的蛋白质组分与人类脑突触膜谷氨酸结合蛋白具有显著的免疫化学相似性。
