Inhibition of growth of human TE2 and C-33A cells by the cell-permeant calpain inhibitor benzyloxycarbonyl-Leu-Leu-Tyr diazomethyl ketone.
Inhibition of growth of human TE2 and C-33A cells by the cell-permeant calpain inhibitor benzyloxycarbonyl-Leu-Leu-Tyr diazomethyl ketone.
复制标题
细胞渗透性钙蛋白酶抑制剂苯甲氧基羰基-Leu-Leu-Tyr 重氮甲基酮抑制人 TE2 和 C-33A 细胞的生长。
DOI：
10.1006/excr.1994.1328
复制
发表时间：
1994
影响因子：
3.7
通讯作者：
Mericle,MT
中科院分区：
文献类型：
--
作者：
Mellgren,RL;Shaw,E;Mericle,MT
钙蛋白酶是一种需要钙离子的非溶酶体蛋白酶，被认为参与细胞内钙离子信号转导的某些方面。然而，其确切的生理功能尚未确定。通过线粒体MTT还原酶活性或直接细胞计数评估，向人TE 2或C-33 A细胞中加入细胞渗透性、不可逆钙蛋白酶抑制剂ZLLY-CHN 2抑制生长。用不含抑制剂的生长培养基替代50 μ MZLLY-CHN 2 24小时暴露产生的生长抑制逆转。ZLLY-CHN 2存在下培养的细胞产生的匀浆显示钙蛋白酶和钙非依赖性蛋白水解活性降低。蛋白质免疫印迹分析表明，细胞培养，失去了80%的钙蛋白酶活性仍然保留完整的钙蛋白酶免疫反应性。因此，ZLLY-CHN 2的抑制似乎导致细胞内不可逆失活的钙蛋白酶的积累。在20或50 μ MZLVG-CHN 2（一种对钙蛋白酶几乎没有活性的细胞渗透性抑制剂）存在下培养的细胞匀浆具有降低的Ca 2+非依赖性蛋白水解活性，但证明钙蛋白酶活性没有降低。在这些条件下，ZLVG-CHN 2不会抑制细胞生长。不表达钙蛋白酶样蛋白酶的酵母细胞的生长不受培养基中含有50 μ MZLLY-CHN 2的抑制。这些结果表明，钙蛋白酶参与多细胞生物细胞生长的社会调节。
Calpains are Ca2+-requiring, nonlysosomal proteases which are thought to participate in some aspects of intracellular Ca2+-signal transduction. However, their exact physiologic function has not yet been established. Addition of the cell-permeant, irreversible calpain inhibitor, ZLLY-CHN2, to human TE2 or C-33A cells inhibited growth, as assessed either by mitochondrial MTT reductase activity or by direct cell counting. Inhibition of growth produced by a 24-h exposure to 50 μMZLLY-CHN2was reversed upon substituting growth medium without inhibitor. Homogenates produced from cells cultured in the presence of ZLLY-CHN2displayed decreased calpain and Ca2+-independent proteolytic activities. Protein immunoblot analysis showed that cell cultures which had lost 80% of their calpain activity still retained full calpain immunoreactivity. Therefore, inhibition by ZLLY-CHN2appeared to result in accumulation of irreversibly inactivated calpain within the cells. Homogenates from cells cultured in the presence of 20 or 50 μMZLVG-CHN2, a cell-permeant inhibitor with little activity against calpains, had decreased Ca2+-independent proteolytic activity, but demonstrated no decrease in calpain activity. ZLVG-CHN2did not inhibit cell growth under these conditions. Growth ofSaccharomyces cerevisiaecells, which do not appear to express calpain-like proteases, was not inhibited by including 50 μMZLLY-CHN2in the culture medium. These results indicate that calpains participate in the social regulation of cell growth in multicellular organisms.