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固定细胞与活细胞之间的细胞相互作用。放射性物质从活细胞向固定细胞的转移。

Cellular interaction between fixed and living cells. Transfer of radioactive materials from living cells to fixed cells.

作者信息

Sakiyama H, Otsu H, Kanegasaki S

出版信息

J Cell Biol. 1979 Jun;81(3):498-509. doi: 10.1083/jcb.81.3.498.

Abstract

Transfer of radioactive materials to fixed cells from an overlying layer of living cells has been examined to determine whether fixed cells can act as acceptors of glycosyltransferases of living cells. After the incubation of living cells were removed by EDTA treatment, and the radioactivity associated with the fixed cells was determined. Lipids, proteins and carbohydrates were found to be transfered from the living cells to the fixed cells. The amount of radioactivity transferred to the fixed cells was dependent on the number of both fixed and living cells and increased with the time of incubation. When fixed cells were treated with chloroform-methanol before the addition of living cells, the transfer of both lipids and proteins to the fixed cells decreased drastically, but only a slight decrease incarbohydrate transfer was observed. Most of the radioactive materials transferred from living cells labeled with glucosamine or fucose to chloroform-methanol-treated fixed cells were solubilized by trypsin but not by the detergents tested. Approximately 55% of the materials transferred from the cells labeled with glucosamine could be solubilized by hyaluronidase and chondroitinase, and the rest was solubilized by neuraminidase and a glycosidase mixture. The treatment of chloroform-methanol-extracted fixed cells with trypsin caused a significant decrease in the transfer from cells labeled with glucosamine. When nucleotide sugars were used as the radioactive precursor, no significant amount of radioactivity was transferred to the fixed cells.

摘要

已对放射性物质从活细胞覆盖层向固定细胞的转移进行了研究,以确定固定细胞是否可作为活细胞糖基转移酶的受体。经乙二胺四乙酸(EDTA)处理去除活细胞后,测定与固定细胞相关的放射性。发现脂质、蛋白质和碳水化合物从活细胞转移到了固定细胞。转移到固定细胞的放射性量取决于固定细胞和活细胞的数量,并随孵育时间增加。在加入活细胞之前,用氯仿 - 甲醇处理固定细胞,脂质和蛋白质向固定细胞的转移大幅减少,但碳水化合物转移仅略有减少。从用氨基葡萄糖或岩藻糖标记的活细胞转移到经氯仿 - 甲醇处理的固定细胞的大多数放射性物质可被胰蛋白酶溶解,但不能被测试的去污剂溶解。从用氨基葡萄糖标记的细胞转移的物质中,约55%可被透明质酸酶和软骨素酶溶解,其余的被神经氨酸酶和糖苷酶混合物溶解。用胰蛋白酶处理经氯仿 - 甲醇提取的固定细胞,会使从用氨基葡萄糖标记的细胞的转移显著减少。当使用核苷酸糖作为放射性前体时,没有大量放射性转移到固定细胞。

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