Fig. 4: NIM directly targets NLRP3’s Lys565 site in the NLRP3 NACHT domain to inhibit oligomerization. | Communications Biology

Fig. 4: NIM directly targets NLRP3’s Lys565 site in the NLRP3 NACHT domain to inhibit oligomerization.

From: Nimbolide ameliorates ARDS and ulcerative colitis by disrupting NLRP3 inflammasome activation

Fig. 4: NIM directly targets NLRP3’s Lys565 site in the NLRP3 NACHT domain to inhibit oligomerization.The alternative text for this image may have been generated using AI.

A DARTS assay performed in BMDMs lysates. Lysates were incubated with NIM or vehicle followed by pronase digestion. B DARTS assay in HEK293T cells transfected with plasmids encoding Flag-NLRP3, Flag-NEK7, or Flag-ASC. C Domain mapping using DARTS in HEK293T cells expressing Flag-tagged LRR, NACHT, or PYD domains of NLRP3. D SPR analysis of the binding affinity between NIM and recombinant human NLRP3 protein. E Analysis of NLRP3 oligomerization by SDD-AGE and total NLRP3 expression by SDS-PAGE in LPS + ATP-stimulated BMDMs treated with NIM. F Molecular docking simulation of NIM (orange, stick) binding to the NLRP3 NACHT domain (cartoon, PDB ID: 6NPY). The enlarged view illustrates the predicted hydrogen bond (yellow dashed line) between NIM and the Lys565 residue (purple sticks). G DARTS assay in HEK293T cells transfected with K565A mutant. Data are representative of three biologically independent experiments.

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