Sequential activation of three distinct ICE‐like activities in Fas‐ligated Jurkat cells

作者：Eric L. Greidinger, Douglas K. Miller, Ting-Ting Yamin, Livia A Casciola-Rosen, Antony Rosen · 发表于：FEBS Letters · 年份：1996 · DOI：10.1016/0014-5793(96)00678-3 · 被引用次数：117 · 研究领域：Cell death mechanisms and regulation、Cell Adhesion Molecules Research、Phagocytosis and Immune Regulation

ICE family proteases have been implicated as important effectors of the apoptotic pathway, perhaps acting hierarchically in a protease cascade. Using cleavage of endogenous protease substrates as probes, three distinct tiers of ICE-like activity were observed after Fas ligation in Jurkat cells. The earliest cleavage detected (30 min) was of fodrin, and produced a 150 kDa fragment. The second phase of cleavage (50 min) involved PARP, U1-70kDa and DNA-PKcs, all substrates of the CPP32-like proteases. Lamin B cleavage was observed during the third cleavage phase (90 min). Distinct inhibition profiles obtained using a panel of peptide-based inhibitors of ICE-like proteases clearly distinguished the three different cleavage phases. These studies provide evidence for a sequence of ICE-like proteolytic activity during apoptosis. The early fodrin cleavage, producing a 150 kDa fragment, identifies an ICE-like activity proximal to CPP32 in Fas-induced Jurkat cell apoptosis.