The pro-form of caspase-8 prevents necroptosis by functioning in a proteolytically active complex with its catalytic-dead homolog, FLICE (FADD [Fas-associated death domain]-like interleukin 1β-converting enzyme)-like inhibitory protein long-form (FLIPL). However, how FLIPL imparts caspase-8 the catalytic activity to suppress necroptosis remains elusive. Here, we show that the protease-like domain of FLIPL is essential for the activity of the caspase-8-FLIPL heterodimer in blocking necroptosis. While substitution of two amino acids whose difference may contribute to the pseudo-caspase property of FLIPL with the corresponding amino acids in caspase-8 does not restore the protease activity of FLIPL, one of the amino acid replacements, tyrosine (Y) 362 to cysteine (C), is sufficient to completely abolish the activity of the caspase-8-FLIPL heterodimer in cleaving receptor-interacting protein 1 (RIP1), thus releasing the necroptosis blockade. Unconstrained necroptosis is observed in embryonic day (E)10.5-E11.5 embryos of FLIPL-Y362C knockin mice. Collectively, these results reveal that the protease-like domain of FLIPL has a special structure that imparts the pro-caspase-8-FLIPL heterodimer a unique catalytic activity toward RIP1 to prevent necroptosis.
Keywords: CP: Molecular biology; FLIP(L); TNF; caspase-8; necroptosis; protease activity.
Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.