nlrp3 protein  (BPS Bioscience)

Journal: Journal of Nanobiotechnology

Article Title: A living therapeutic platform for localized in situ modulation of macrophage pyroptosis ameliorates GVHD while preserving GVL

doi: 10.1186/s12951-026-04285-6

Figure Lengend Snippet: Schematic of the engineered probiotic strategy for treating intestinal GVHD. Orally administered, engineered Escherichia coli Nissle 1917 produces outer membrane vesicles (OMVs) displaying the NLRP3-inhibitory peptide BigLEN in the gut. These OMVs enable delivery of the displayed peptide to intestinal tissues and are uptake by lamina propria macrophages. Inside macrophages, BigLEN binds to the NLRP3 NACHT domain, inhibiting inflammasome assembly and subsequent pyroptosis. By attenuating macrophage pyroptosis and the associated cytokine storm, the treatment reduces the activation and infiltration of pathogenic T‑cell subsets (such as Th1 and Th17 cells) and promotes a shift in macrophage polarization toward an anti‑inflammatory phenotype. Ultimately, this locally restricted immunomodulation alleviates intestinal GVHD

Article Snippet: Surface plasmon resonance (SPR) analysis was performed on a Biacore T200 instrument (GE Healthcare, USA) to measure the binding affinity between the NACHT protein (HY-P701027, MCE, USA) and BigLEN (501036-69-7, MCE, USA).

Techniques: Membrane, Activation Assay

Journal: Journal of Nanobiotechnology

Article Title: A living therapeutic platform for localized in situ modulation of macrophage pyroptosis ameliorates GVHD while preserving GVL

doi: 10.1186/s12951-026-04285-6

Figure Lengend Snippet: Identification and functional validation of BigLEN as a novel NLRP3‑inhibitory peptide. ( A ) Schematic workflow for the virtual screening of NLRP3‑inhibitory peptides. ( B ) Structural analysis of the NLRP3 NACHT domain in complex with a known inhibitor, highlighting the conserved hydrophobic binding pocket. ( C ) Pharmacophore model constructed based on key inhibitor‑protein interactions, featuring four critical chemical features (hydrogen bond acceptors and donors) for virtual screening. ( D ) Molecular docking pose of the candidate peptide BigLEN within the NACHT domain hydrophobic pocket. ( E ) Detailed interaction diagram between BigLEN and key residues (His367, Arg578, Glu369, Glu629) of the NLRP3 NACHT domain, with corresponding binding energies. ( F ) Surface plasmon resonance (SPR) sensograms showing concentration‑dependent binding of BigLEN to immobilized NLRP3 protein. The equilibrium dissociation constant (KD) was calculated to be 1.749 × 10⁻⁷ M. ( G - H ) Flow cytometry analysis of cell death in mouse bone marrow‑derived macrophages (BMDMs). Cells were primed with LPS and stimulated with nigericin in the presence or absence of BigLEN (10 or 50 µM). ( I ) Representative Western blot images of key pyroptosis‑related proteins and ASC in BMDMs in different groups. ( J ) Lactate dehydrogenase (LDH) release assay of supernatant from BMDMs following LPS/nigericin stimulation in the presence or absence of BigLEN (10 or 50 µM), supernatants were collected 12 h after BigLEN treatment. ( K ) Representative TEM image of BMDMs in different groups. Scale bar = 1 μm. Data are representative of three independent experiments. Statistical significance was determined by one‑way ANOVA with Tukey’s post‑hoc test (* p < 0.05). Quantitative data are presented as mean ± SEM

Article Snippet: Surface plasmon resonance (SPR) analysis was performed on a Biacore T200 instrument (GE Healthcare, USA) to measure the binding affinity between the NACHT protein (HY-P701027, MCE, USA) and BigLEN (501036-69-7, MCE, USA).

Techniques: Functional Assay, Biomarker Discovery, Binding Assay, Construct, SPR Assay, Flow Cytometry, Western Blot, Lactate Dehydrogenase Assay

Journal: Pharmaceuticals

Article Title: In Silico and In Vitro Evaluation of Quercetin Metabolites Binding to Inflammatory Target Proteins

doi: 10.3390/ph19050655

Figure Lengend Snippet: Molecular docking analysis. ( A ) The tested compounds (quercetin, quercetin 3- O -glucuronide, quercetin 7- O -glucuronide, isorhamnetin, isorhamnetin 3- O -glucuronide, and the positive control (MCC950) were inside the binding site of the NLRP3 (pdb:6npy) protein. ( B ) Quercetin, isorhamnetin, and MCC950 with the interacting amino acids.

Article Snippet: Recombinant NLRP3 (Origene, TP750176) was labeled with the Monolith Protein Labeling Kit RED-NHS 2nd Generation (Cat. No. M0-L011, NanoTemper Technologies, Munich, Germany).

Techniques: Positive Control, Binding Assay

Journal: Pharmaceuticals

Article Title: In Silico and In Vitro Evaluation of Quercetin Metabolites Binding to Inflammatory Target Proteins

doi: 10.3390/ph19050655

Figure Lengend Snippet: Binding of quercetin, isorhamnetin, quercetin 7- O -glucuronide, quercetin 3- O -glucuronide, isorhamnetin 3- O -glucuronide, tamarixetin, hippuric acid, and 3,4-dihydroxytoluene to NLRP3 as determined by microscale thermophoresis.

Article Snippet: Recombinant NLRP3 (Origene, TP750176) was labeled with the Monolith Protein Labeling Kit RED-NHS 2nd Generation (Cat. No. M0-L011, NanoTemper Technologies, Munich, Germany).

Techniques: Binding Assay, Microscale Thermophoresis