Yang Chen, Wang Ling, Liu Yuchen, Zhang Yuehui, Jin Chaozhi, Cheng Jiale, Shang Limin, Fang Longlong, Wu Shanshan, Chen Chuan, Wang Jian
College of Life Sciences, Hebei University, Baoding, China; State Key Laboratory of Medical Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing, China.
State Key Laboratory of Medical Proteomics, Beijing Proteome Research Center, National Center for Protein Sciences (Beijing), Beijing Institute of Lifeomics, Beijing, China.
Mol Cell Proteomics. 2025 May;24(5):100972. doi: 10.1016/j.mcpro.2025.100972. Epub 2025 Apr 16.
NOD-like receptor (NLR) family pyrin domain containing 3 (NLRP3) involves in inflammasome complex assembly and innate immunity. Activation of the NLRP3 inflammasome induces conformational alterations in protein complexes, influencing their interactions with other molecules, which in turn affects protein thermal stability. To investigate the proteome-wide thermal stability alterations induced by NLRP3 inflammasome activation, we conducted a comprehensive analysis of meltome dynamics using thermal proteome profiling. Our analysis identified 337 proteins exhibiting alterations in thermal stability upon NLRP3 inflammasome activation. Subsequently, we validated three proteins by the cellular thermal shift assay. Notably, our findings reveal that the majority of these proteins tend to cluster into distinct macromolecular complexes. Furthermore, we identified FAM120A as a novel NLRP3 binding partner, with its suppression enhancing caspase-1 activation and IL-1β release in response to NLRP3 agonist. Collectively, these data provide a comprehensive framework for understanding the mechanisms of NLRP3 inflammasome activation and underscore the utility of thermal proteome profiling in exploring proteome-wide thermal stability changes during signaling transduction.
含NOD样受体(NLR)家族pyrin结构域3(NLRP3)参与炎性小体复合物组装和固有免疫。NLRP3炎性小体的激活诱导蛋白质复合物的构象改变,影响它们与其他分子的相互作用,进而影响蛋白质的热稳定性。为了研究NLRP3炎性小体激活诱导的全蛋白质组热稳定性改变,我们使用热蛋白质组分析对熔解组动力学进行了全面分析。我们的分析鉴定出337种蛋白质在NLRP3炎性小体激活后热稳定性发生改变。随后,我们通过细胞热位移分析验证了三种蛋白质。值得注意的是,我们的研究结果表明,这些蛋白质中的大多数倾向于聚集成不同的大分子复合物。此外,我们鉴定出FAM120A是一种新型的NLRP3结合伴侣,抑制它可增强对NLRP3激动剂的半胱天冬酶-1激活和白细胞介素-1β释放。总的来说,这些数据为理解NLRP3炎性小体激活机制提供了一个全面的框架,并强调了热蛋白质组分析在探索信号转导过程中全蛋白质组热稳定性变化方面的实用性。