Gasdermin D (GSDMD) is the chief executioner of inflammatory cell death or pyroptosis. During pyroptosis, proteolytic processing of GSDMD releases its N-terminal domain (NTD), which then forms large oligomeric pores in the plasma membranes. Membrane pore-formation by NTD allows the release of inflammatory cytokines and causes membrane damage to induce cell death. Structural mechanisms of GSDMD-mediated membrane pore-formation have been extensively studied. However, less effort has been made to understand the physicochemical properties of GSDMD and their functional implications. Here, we explore detailed characterization of the physicochemical properties of mouse GSDMD (mGSDMD), and their implications in regulating the pore-forming function. Our study reveals that mGSDMD shows some of the hallmark features of amyloids, and forms oligomeric assemblies in solution that are critically dependent on the disulfide bond-forming ability of the protein. mGSDMD oligomeric assemblies do not resemble typical amyloid fibrils/aggregates, and do not show resistance to proteolytic degradation that is otherwise observed with the conventional amyloids. Our results further elucidate the essential role of an amyloid-prone region (APR) in the oligomerization and amyloid-like features of mGSDMD. Furthermore, alteration of this APR leads to compromised pore-forming ability and cell-killing activity of NTD released from mGSDMD. Taken together, our study for the first time provides crucial new insights regarding implications of the amyloid-like property of mGSDMD in regulating its pore-forming function, which is an essential requirement for this pyroptotic executioner. To the best of our knowledge, such mode of regulation of mGSDMD-function has not been appreciated so far.

中文翻译：

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焦亡执行者成孔蛋白 gasdermin D 形成寡聚组装并表现出淀粉样蛋白特性，可能有助于其造孔功能。


Gasdermin D （GSDMD） 是炎性细胞死亡或细胞焦亡的主要执行者。在焦亡过程中，GSDMD 的蛋白水解加工释放其 N 末端结构域 （NTD），然后在质膜中形成大的寡聚孔。NTD 形成膜孔允许炎性细胞因子释放并导致膜损伤以诱导细胞死亡。GSDMD 介导的膜孔形成的结构机制已被广泛研究。然而，在了解 GSDMD 的物理化学性质及其功能意义方面所做的努力较少。在这里，我们探讨了小鼠 GSDMD （mGSDMD） 的物理化学性质的详细表征，以及它们在调节成孔功能中的意义。我们的研究表明，mGSDMD 显示出淀粉样蛋白的一些标志性特征，并在溶液中形成寡聚组装体，这些组装体严重依赖于蛋白质的二硫键形成能力。mGSDMD 寡聚组装体与典型的淀粉样蛋白原纤维/聚集体不同，并且不表现出对传统淀粉样蛋白所观察到的蛋白水解降解的抵抗力。我们的结果进一步阐明了淀粉样蛋白易感区域 （APR） 在 mGSDMD 的寡聚化和淀粉样蛋白特征中的重要作用。此外，这种 APR 的改变会导致 mGSDMD 释放的 NTD 的成孔能力和细胞杀伤活性受损。综上所述，我们的研究首次提供了关于 mGSDMD 的淀粉样蛋白特性在调节其成孔功能中的影响的重要新见解，这是这种焦亡执行者的基本要求。据我们所知，到目前为止，这种 mGSDMD 功能的调节模式尚未得到重视。