Keuler Tim, Gatterdam Karl, Akbal Anil, Lovotti Marta, Marleaux Michael, Geyer Matthias, Latz Eicke, Gütschow Michael
Pharmaceutical Institute, University of Bonn, Bonn, Germany.
Institute of Structural Biology, University of Bonn, Bonn, Germany.
Front Chem. 2021 Apr 22;9:642273. doi: 10.3389/fchem.2021.642273. eCollection 2021.
Extracellular signals drive the nucleation of the NLRP3 inflammasome which leads to the release of cytokines and causes inflammatory events. Hence, the inflammasome has gained enormous momentum in biomedical basic research. The detailed mechanisms of inflammasome generation and regulation remain to be elucidated. Our study was directed toward the design, convergent synthesis, and initial biochemical evaluation of activity-based probes addressing NLRP3. For this purpose, probes were assembled from a CRID3/MCC950-related NLRP3-binding unit, a linker portion and a coumarin 343 fluorophore or biotin. The affinity of our probes to NLRP3 was demonstrated through SPR measurements and their cellular activity was confirmed by reduction of the interleukin 1β release from stimulated bone marrow-derived macrophages. The initial characterizations of NLRP3-targeting probes highlighted the coumarin probe as a suitable tool compound for the cellular and biochemical analysis of the NLRP3 inflammasome.
细胞外信号驱动NLRP3炎性小体的成核，这会导致细胞因子的释放并引发炎症事件。因此，炎性小体在生物医学基础研究中获得了巨大的发展动力。炎性小体产生和调节的详细机制仍有待阐明。我们的研究旨在设计、汇聚合成并初步生化评估针对NLRP3的基于活性的探针。为此，探针由一个与CRID3/MCC950相关的NLRP3结合单元、一个连接部分和一个香豆素343荧光团或生物素组装而成。通过表面等离子体共振测量证明了我们的探针与NLRP3的亲和力，并通过减少刺激的骨髓来源巨噬细胞中白细胞介素1β的释放证实了它们的细胞活性。靶向NLRP3探针的初步表征突出了香豆素探针作为用于NLRP3炎性小体细胞和生化分析的合适工具化合物。
