Fig. 3: PQS binds the intracellular receptor CNMT, identified via MS-CETSA profiling, to promote its enzymatic activities. | Nature Communications

Fig. 3: PQS binds the intracellular receptor CNMT, identified via MS-CETSA profiling, to promote its enzymatic activities.

From: A Pseudomonas aeruginosa quorum-sensing metabolite manipulates macrophage ferroptosis through a methylation pathway

Fig. 3: PQS binds the intracellular receptor CNMT, identified via MS-CETSA profiling, to promote its enzymatic activities.

A MS-CETSA experimental procedure to identify the potential direct target of PQS. Schematic diagrams were created using BioRender.com. B Volcano plot of T-test screening of MS-CETSA assay results to identify CNMT as the potential direct target of PQS. Data are presented as the log2 fold change in protein abundance (x-axis) versus the -log10 p-value (y-axis) for each protein, derived from triplicate experiments. Two-sided moderated t test p-values were calculated. C Representative western blot analysis of CETSA experiments of RAW264.7 cell lysate in the presence of PQS or DMSO under 37, 46, 49, 52, 55, and 58 °C. Thermally stable Superoxide Dismutase 1 (SOD1) served as a loading control. D The interaction model of the CNMT protein with PQS visualised using Chimera. E Interaction map between residues of the CNMT protein with PQS. F Quantifying the binding affinity of CNMT protein with PQS using MST assay (n = 3). G Steady-state kinetics of CNMT and CNMT + PQS for carnosine. Kinetic measurements were performed using a bioluminescence methyltransferase assay, MTase-Glo (Promega). Data are presented as mean ± SD (n = 3, two-way ANOVA with Tukey’s multiple comparisons test). Source data are provided as a Source Data file.

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