Kornilova E S, Bobrova I F, Nesterov A M, Nikol'skiĭ N N, Sorokin A B
Tsitologiia. 1988 Mar;30(3):291-8.
The compartmentalization of the epidermal growth factor (EGF) receptors in A-431 cells was studied using centrifugation of the microsomal fraction of these cells in continuous Percoll gradient. The existence of an intact (non-degraded) EGF receptor in plasma membrane and endosome fraction was demonstrated by electrophoretic analysis of in vitro phosphorylated Percoll fractions. No phosphorylated receptor was revealed in lysosomal fraction by this method. The existence of non dissociated EGF-receptor complexes in intracellular compartments 30 minutes after the start of internalization was proven using a synthesized photoreactive labeled EGF derivative (125I-EGF-SANAH). The removing of pH gradient in organellar membranes by 10 mkM of monensin did not affect dissociation from its receptor. The data obtained proved the existence of non-dissociated and non-degraded EGF-receptor complexes in the endosomal compartment of A-431 cells.
利用这些细胞微粒体部分在连续Percoll梯度中的离心作用,对A - 431细胞中表皮生长因子(EGF)受体的区室化进行了研究。通过对体外磷酸化的Percoll组分进行电泳分析,证实了质膜和内体组分中存在完整(未降解)的EGF受体。用这种方法在溶酶体组分中未发现磷酸化受体。使用合成的光反应性标记EGF衍生物(125I - EGF - SANAH),证明了内化开始30分钟后细胞内区室中存在未解离的EGF - 受体复合物。10 mkM莫能菌素去除细胞器膜中的pH梯度并不影响其与受体的解离。所获得的数据证明了A - 431细胞内体区室中存在未解离和未降解的EGF - 受体复合物。