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首页> 外文期刊>Molecular cancer therapeutics >Modifications enhance the apoptosis-inducing activity of FADD.
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Modifications enhance the apoptosis-inducing activity of FADD.

机译:修饰增强了FADD的凋亡诱导活性。

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The ability to enhance apoptosis-inducing activity in specific cells, despite the presence of cellular antiapoptotic proteins, would allow the removal of target cells from a cell population. Here, we show that modification of Fas-associated protein with death domain (FADD) by fusing the tandem death effector domains (DED) of FADD to the E protein of lambda phage, a head coat protein with self-assembly activity, greatly increases the apoptosis-inducing activity of FADD in both adherent NIH3T3 and HEK293 cells. Induction of apoptosis in cell lines that stably express modified FADD (2DEDplusE) resulted in rapid blebbing, and most cells detached from the flask within 5 h. In contrast, following induction of apoptosis, it took over 24 h for the cells expressing unmodified FADD to exhibit these signs. The cells expressing the modified FADD underwent apoptosis through the typical apoptosis cascade via activation of caspase-3, and apoptosis was inhibited by a caspase inhibitor (i.e., z-VAD-fmk). Theoretically,as our adhesive stable cell lines undergo apoptosis rapidly and in synchrony following mifepristone- or tetracycline-controlled production of a single apoptosis protein without affecting any other cellular pathways, they provide excellent model systems in which to analyze the phenomenon of apoptosis in adhesive cell lines, in particular, blebbing and detachment.
机译:尽管存在细胞抗凋亡蛋白,但增强特定细胞凋亡诱导活性的能力仍可从细胞群中去除靶细胞。在这里,我们表明,通过将FADD的串联死亡效应子域(DED)融合到具有自组装活性的头皮蛋白E蛋白中,可以将Fas相关蛋白与死亡域(FADD)融合在一起。粘附的NIH3T3和HEK293细胞中FADD的凋亡诱导活性。稳定表达修饰的FADD(2DEDplusE)的细胞系中细胞凋亡的诱导导致快速起泡,大多数细胞在5小时内从烧瓶中脱落。相反,诱导细胞凋亡后,表达未修饰的FADD的细胞需要24小时才能显示出这些迹象。表达修饰的FADD的细胞通过激活caspase-3通过典型的凋亡级联反应经历凋亡,并且凋亡被caspase抑制剂(即z-VAD-fmk)抑制。从理论上讲,由于我们的黏附稳定细胞系在米非司酮或四环素控制的单个凋亡蛋白产生后迅速且同步地发生凋亡,而不会影响任何其他细胞途径,因此它们提供了出色的模型系统,可用于分析黏附细胞的凋亡现象线条特别是起泡和脱离。

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