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首页> 外文期刊>Molecular and Cellular Biology >Rapid cell surface appearance of endocytic membrane proteins in Chinese hamster ovary cells.
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Rapid cell surface appearance of endocytic membrane proteins in Chinese hamster ovary cells.

机译：中国仓鼠卵巢细胞内吞膜蛋白的快速细胞表面出现。

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Lactoperoxidase was used to selectively radiolabel endocytic membrane. CHO cells were incubated with enzyme at 37 degrees C for 10 min to permit lactoperoxidase internalization. Radioiodination was done at 4 degrees C. About 90% of the radioiodinated products pelleted at 100,000 X g. From 12 to 15 different electrophoretic species were detected by one-dimensional gel electrophoresis. When cells labeled by internalized lactoperoxidase were warmed to 37 degrees C, the incorporated radioactivity was lost in a biphasic manner with an overall t1/2 of approximately 20 h. Upon warming cells to 37 degrees C, the labeled species became sensitive to pronase or trypsin digestion. The increase in protease sensitivity was progressive over a 10- to 20-min period. Maximally 45% of the initially intracellular radiolabel could be released. A digest of exterior-radioiodinated cells released 50% of the incorporated radioiodine. These observations strongly suggest a rapid shuttling of approximately 90% of the radioiodinated membrane species initially present within the cell to the cell surface.
机译：乳过氧化物酶用于选择性地放射性标记内吞膜。将CHO细胞与酶在37℃下孵育10分钟，以使乳过氧化物酶内在化。放射性碘化在4摄氏度下进行。约90％的放射性碘化产品以100,000 X g沉淀。一维凝胶电泳可检测出12至15种不同的电泳物质。当用内在的乳过氧化物酶标记的细胞加热到37摄氏度时，合并的放射性以两相方式丧失，总t1 / 2约为20小时。将细胞加热到37摄氏度后，标记的物种对链酶或胰蛋白酶消化变得敏感。蛋白酶敏感性的增加在10至20分钟的时间内逐渐进行。最初的细胞内放射性标记最多可释放45％。外部放射性碘化细胞的消化物释放了50％的掺入放射性碘。这些观察结果强烈表明，最初存在于细胞中的放射性碘化膜物质中大约90％会快速穿梭到细胞表面。

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