Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line.
Vectorial targeting of apical and basolateral plasma membrane proteins in a human adenocarcinoma epithelial cell line.
复制标题
DOI：
10.1073/pnas.86.23.9313
复制
发表时间：
1989-12
影响因子：
11.1
通讯作者：
A. Bivic;Francisco X. REALt;Enrique;RODRIGUEZ-BOULAN
中科院分区：
文献类型：
--
作者：
A. Bivic;Francisco X. REALt;Enrique;RODRIGUEZ-BOULAN
我们研究了肠细胞中新合成的质膜蛋白所遵循的表面递送途径。为此，我们开发了一种分析方法，并鉴定了一种来源于人结肠腺癌的上皮细胞系(SK-CO-15)。用放射性蛋氨酸/半胱氨酸冲击生长在聚碳酸酯滤池上的极化融合单层(2000 omega.cm2)，在不同的追逐时间，通过区域选择性生物素化、免疫沉淀和固定化链霉亲和素沉淀回收到达顶端或基底侧面的蛋白质组分。结果表明，根尖标记和根尖标记均以矢量方式传递到各自的表面，分选效率分别为50：1和14：1。
We studied the surface delivery pathways followed by newly synthesized plasma membrane proteins in intestinal cells. To this end, we developed an assay and characterized an epithelial cell line (SK-CO-15) derived from human colon adenocarcinoma. Polarized confluent monolayers (2000 omega.cm2), grown on polycarbonate filter chambers, were pulsed with radioactive methionine/cysteine and, at different times of chase, the protein fraction reaching the apical or basolateral surface was recovered by domain-selective biotinylation, immunoprecipitation, and immobilized streptavidin precipitation. Both an apical and a basolateral marker were found to be delivered vectorially to the respective surface, with a sorting efficiency of 50:1 for the basolateral marker and 14:1 for the apical marker.