Pore-forming activity and structural autoinhibition of the gasdermin family
Pore-forming activity and structural autoinhibition of the gasdermin family
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DOI：
10.1038/nature18590
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发表时间：
2016-07-07
期刊：
影响因子：
64.8
通讯作者：
Shao, Feng
中科院分区：
文献类型：
--
作者：
Ding, Jingjin;Wang, Kun;Shao, Feng
炎症性半胱天冬酶切割gasdermin D（GSDMD）蛋白以触发细胞凋亡，这是一种细胞死亡的裂解形式，对免疫防御和疾病至关重要。GSDMD含有gasdermin家族共有的功能重要的gasdermin-N结构域。gasdermin蛋白的功能作用机制尚不清楚。在这里，我们表明，gasdermin蛋白GSDMD，GSDMA 3和GSDMA的gasdermin-N域可以结合膜脂，磷酸肌醇和心磷脂，并表现出膜破坏细胞毒性在哺乳动物细胞和人工转化的细菌。Gasdermin-N在焦亡期间移动到质膜。纯化gasdermin-N有效地裂解磷脂酰肌醇/心磷脂的脂质体，并形成人工或天然磷脂混合物制成的膜上的孔。大多数gasdermin孔的内径为10-14 nm，含有16个对称的原聚体。GSDMA 3的晶体结构显示出在gasdermin家族中保守的自抑制双结构域结构。结构导向突变表明gasdermin-N结构域的脂质体渗漏和孔形成活性是焦亡所需的。这些发现揭示了焦亡的机制，并提供了深入了解gasdermin家族在坏死，免疫和疾病中的作用。
Inflammatory caspases cleave the gasdermin D (GSDMD) protein to trigger pyroptosis, a lytic form of cell death that is crucial for immune defences and diseases. GSDMD contains a functionally important gasdermin-N domain that is shared in the gasdermin family. The functional mechanism of action of gasdermin proteins is unknown. Here we show that the gasdermin-N domains of the gasdermin proteins GSDMD, GSDMA3 and GSDMA can bind membrane lipids, phosphoinositides and cardiolipin, and exhibit membrane-disrupting cytotoxicity in mammalian cells and artificially transformed bacteria. Gasdermin-N moved to the plasma membrane during pyroptosis. Purified gasdermin-N efficiently lysed phosphoinositide/cardiolipin-containing liposomes and formed pores on membranes made of artificial or natural phospholipid mixtures. Most gasdermin pores had an inner diameter of 10-14 nm and contained 16 symmetric protomers. The crystal structure of GSDMA3 showed an autoinhibited two-domain architecture that is conserved in the gasdermin family. Structure-guided mutagenesis demonstrated that the liposome-leakage and pore-forming activities of the gasdermin-N domain are required for pyroptosis. These findings reveal the mechanism for pyroptosis and provide insights into the roles of the gasdermin family in necrosis, immunity and diseases.