Identification of a novel caspase cleavage motif AEAD  

作　　者：Yujie Fang Zhou Gong Miaomiao You Ke Peng 

机构地区：[1]State Key Laboratory of Virology,Center for Antiviral Research,Center for Biosafety Mega-Science,Wuhan Institute of Virology,Chinese Academy of Sciences,Wuhan,430207,China [2]University of Chinese Academy of Sciences,Beijing,100049,China [3]State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics,Innovation Academy for Precision Measurement Science and Technology Chinese Academy of Sciences,Wuhan,430071,China [4]Provincial Key Laboratory of Jiangxia,Wuhan,430207,China

出　　处：《Virologica Sinica》2024年第5期755-766,共12页中国病毒学（英文版）

基　　金：supported by the National Key R&D Program of China(2021YFC2300700);National Science and Technology Major Project(No.2018ZX10101004001005);National Natural Science Foundation of China(numbers 32070179).We thank Dr.Qinxue Hu(Wuhan Institute of Virology)and Dr.Yuchen Xia(Wuhan University)for help with materials.We thank Ding Gao from Center for Instrumental Analysis and Metrology at Wuhan Institute of Virology for his help with the Leica confocal microscope and the Operetta.

摘　　要：Infections of many viruses induce caspase activation to regulate multiple cellular pathways,including programmed cell death,immune signaling and etc.Characterizations of caspase cleavage sites and substrates are important for understanding the regulation mechanisms of caspase activation.Here,we identified and analyzed a novel caspase cleavage motif AEAD,and confirmed its caspase dependent cleavage activity in natural substrate,such as nitric oxide-associated protein 1(NOA1).Fusing the enhanced green fluorescent protein(EGFP)with the mitochondrial marker protein Tom20 through the AEAD motif peptide localized EGFP to the mitochondria.Upon the activation of caspase triggered by Sendai virus(SeV)or herpes simplex virus type 1(HSV-1)infection,EGFP diffusely localized to the cell due to the caspase-mediated cleavage,thus allowing visual detection of the virusinduced caspase activation.An AEAD peptide-derived inhibitor Z-AEAD-FMK were developed,which significantly inhibited the activities of caspases-1,-3,-6,-7,-8 and-9,exhibiting a broad caspase inhibition effect.The inhibitor further prevented caspases-mediated cleavage of downstream substrates,including BID,PARP1,LMNA,pro-IL-1β,pro-IL-18,GSDMD and GSDME,protecting cells from virus-induced apoptotic and pyroptotic cell death.Together,our findings provide a new perspective for the identification of novel caspase cleavage motifs and the development of new caspase inhibitors and anti-inflammatory drugs.

关 键 词：CASPASE AEAD motif INHIBITOR VIRUS Cell death 

分 类 号：S852.65[农业科学—基础兽医学]