Extracellular ATP reversibly increases the plasma membrane permeability of transformed mouse fibroblasts to large macromolecules.
Extracellular ATP reversibly increases the plasma membrane permeability of transformed mouse fibroblasts to large macromolecules.
复制标题
细胞外 ATP 可逆地增加转化小鼠成纤维细胞的质膜对大分子的通透性。
DOI：
10.1006/abio.1993.1080
复制
发表时间：
1993
影响因子：
2.9
通讯作者：
Weisman,GA
中科院分区：
文献类型：
--
作者：
Saribaş,AS;Lustig,KD;Zhang,X;Weisman,GA
细胞外ATP增加转化小鼠成纤维细胞（3 T6细胞）的质膜对异硫氰酸荧光素-葡聚糖（4和9 kDa）和[3 H]菊粉（5.2 kDa）的渗透性。在加入ATP后20至30分钟，对大分子的渗透性增加，随后对小分子（<1 kDa）的渗透性增加。小分子和大分子的渗透性变化以相同的方式通过pH值、温度和ATP浓度进行调节。同样，之前用于在ATP处理后重新密封3 T6细胞的条件使得分子<1 kDa能够进入，也重新密封了用ATP处理20至30分钟的细胞。重新密封的细胞以与对照细胞相同的速率繁殖，尽管许多细胞从培养皿中分离。水母发光蛋白，一个20 kDa的Ca 2+敏感的光蛋白，密封内ATP处理的3 T6细胞，然后用于定量细胞质游离Ca 2+浓度的变化。ATP和离子霉素，钙离子载体，增加水母发光蛋白在3 T6细胞，表明细胞质游离钙水平的升高。摄取大分子（4-20 kDa）的时间过程与用ATP处理的3 T6细胞释放乳酸脱氢酶（150 kDa）的时间过程相似，表明该方法通常可用于将高分子量大分子掺入活的ATP敏感性细胞中。
Extracellular ATP increases the plasma membrane permeability of transformed mouse fibroblasts (3T6 cells) to fluorescein isothiocyanate-dextrans (4 and 9 kDa) and [3H]inulin (5.2 kDa). This increase in permeability to large macromolecules occurred 20 to 30 min after ATP addition, subsequent to an increase in permeability to small macromolecules (<1 kDa). Permeability changes to small and large macromolecules were modulated in the same manner by pH, temperature, and ATP concentration. Likewise, conditions previously used to reseal 3T6 cells after ATP treatments that enabled the entry of molecules <1 kDa also resealed cells that had been treated with ATP for 20 to 30 min. Resealed cells propagated at the same rate as control cells, although many cells became detached from the culture dish. Aequorin, a 20-kDa Ca2+-sensitive photoprotein, was sealed inside ATP-treated 3T6 cells and then used to quantitate changes in the concentration of cytoplasmic free Ca2+. Both ATP and ionomycin, a calcium ionophore, increased aequorin luminescence in 3T6 cells, indicative of an elevation in levels of cytoplasmic free Ca2+. The time course for uptake of macromolecules (4-20 kDa) was similar to the time course of lactic dehydrogenase (150 kDa) release from 3T6 cells treated with ATP, suggesting that the procedure may be generally applicable for incorporation of high molecular weight macromolecules into viable, ATP-sensitive cells.