Multicomponent protein complexes called inflammasomes play a major role in the innate immune system by activating proinflammatory cytokines and promoting a highly inflammatory form of programmed cell death, called pyroptosis. A hallmark of the function of the nucleotide-binding domain, leucine-rich repeat and NLRP3-mediated inflammasome assembly is the polymerization of ASC into large filaments. The ASC filaments recruit and activate procaspase-1 by induced proximity. We developed an in vitro assay for monitoring the polymerization of the pyrin domain of ASC by microscale thermophoresis. We have validated the assay by analyzing the effects of buffer conditions, mutations of ASC and the use of seeds on the polymerization behavior of ASC.

中文翻译：

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用于检测 ASC pyrin 结构域聚合的体外试验的开发。

称为炎症小体的多组分蛋白质复合物通过激活促炎细胞因子和促进高度炎症形式的程序性细胞死亡（称为细胞焦亡）在先天免疫系统中发挥重要作用。核苷酸结合域、富含亮氨酸的重复序列和 NLRP3 介导的炎性体组装的功能的标志是 ASC 聚合成大丝。ASC 细丝通过诱导接近来募集和激活 procaspase-1。我们开发了一种体外检测方法，用于通过微型热泳监测 ASC pyrin 结构域的聚合。我们通过分析缓冲条件、ASC 突变和使用种子对 ASC 聚合行为的影响来验证该测定。