A FRET biosensor for necroptosis uncovers two different modes of the release of DAMPs.
A FRET biosensor for necroptosis uncovers two different modes of the release of DAMPs.
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DOI：
10.1038/s41467-018-06985-6
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发表时间：
2018-10-26
影响因子：
16.6
通讯作者：
Nakano H
中科院分区：
文献类型：
--
作者：
Murai S;Yamaguchi Y;Shirasaki Y;Yamagishi M;Shindo R;Hildebrand JM;Miura R;Nakabayashi O;Totsuka M;Tomida T;Adachi-Akahane S;Uemura S;Silke J;Yagita H;Miura M;Nakano H
坏死性下垂是一种受调节的坏死形式，依赖于受体相互作用蛋白激酶(RIPK)3和混合谱系激酶域样蛋白(MLKL)。虽然危险相关分子模式(DAMP)S参与了各种病理条件，并从死亡细胞中释放出来，但其潜在的机制尚不完全清楚。在这里，我们开发了一种荧光共振能量转移(FRET)生物传感器，称为SMART(一种基于FRET的RIPK3激活MLKL的传感器)。SMART由MLKL片段组成，监测坏死性下垂，但不监测细胞凋亡或坏死。从机制上讲，SMART监测由RIPK3或突变激活诱导的寡聚化MLKL的质膜转位。SMART结合核湿释放成像和分泌活性活细胞成像(LCI-S)揭示了从坏死性细胞释放高迁移率族蛋白1的两种不同模式。因此，SMART和LCI-S发现了坏死性下垂过程中湿气释放的新规律。趋化细胞激活MLKL并释放炎症介质，尽管这一过程的潜在调控机制尚不清楚。在这里，Murai等人。开发一种坏死下垂特异性FRET传感器(SMART)，监测MLKL膜转位，以识别两种湿气释放模式。
Necroptosis is a regulated form of necrosis that depends on receptor-interacting protein kinase (RIPK)3 and mixed lineage kinase domain-like (MLKL). While danger-associated molecular pattern (DAMP)s are involved in various pathological conditions and released from dead cells, the underlying mechanisms are not fully understood. Here we develop a fluorescence resonance energy transfer (FRET) biosensor, termed SMART (a sensor for MLKL activation by RIPK3 based on FRET). SMART is composed of a fragment of MLKL and monitors necroptosis, but not apoptosis or necrosis. Mechanistically, SMART monitors plasma membrane translocation of oligomerized MLKL, which is induced by RIPK3 or mutational activation. SMART in combination with imaging of the release of nuclear DAMPs and Live-Cell Imaging for Secretion activity (LCI-S) reveals two different modes of the release of High Mobility Group Box 1 from necroptotic cells. Thus, SMART and LCI-S uncover novel regulation of the release of DAMPs during necroptosis. Necroptotic cells activate MLKL and release inflammatory DAMPs, although the underlying regulatory mechanisms of this process are poorly understood. Here, Murai et al. develop a necroptosis-specific FRET sensor (SMART) that monitors MLKL membrane translocation to identify two modes of DAMP release.
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影响因子：
16
作者：
Quarato G;Guy CS;Grace CR;Llambi F;Nourse A;Rodriguez DA;Wakefield R;Frase S;Moldoveanu T;Green DR
通讯作者：
Green DR
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46.9
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6.7
作者：
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通讯作者：
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影响因子：
64.5
作者：
Gong YN;Guy C;Olauson H;Becker JU;Yang M;Fitzgerald P;Linkermann A;Green DR
通讯作者：
Green DR