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

FEBS Letters

Rosen, Antony¹  Yamin, Ting-Ting³  Greidinger, Eric L.¹  Casciola-Rosen, Livia²  Miller, Douglas K.³ 

[1] Department of Medicine, Johns Hopkins University School of Medicine, 720 Rutland Avenue, Room 1055, Baltimore, MD 21205, USA;Department of Dermatology, Johns Hopkins University School of Medicine, 720 Rutland Avenue, Baltimore, MD 21205, USA;Department of Biochemistry and Inflammation Research, Merck Research Laboratories, P.O. Box 2000, Rahway, NJ 07065, USA

关键词: Apoptosis;    Protease;    CPP32;    ICE;   

DOI  :  10.1016/0014-5793(96)00678-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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-PK_cs, 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.

【 授权许可】

Unknown