Figure 5.

TH6744 treatment alters host proteostasis pathways. (A) Visualization of known and predicted protein–protein interaction network between 79 overlapping significant proteins in TPP-TR and 2D-TPP analysis by STRING database and Cytoscape tool with minimum required interaction score 0.7 (high confidence). Each node represents a protein, line represents an experimentally proven or predicted interaction, node size represents ΔTm between TH6744 and DMSO-treated sample from TPP-TR and node color represents stabilization (red) or destabilization (yellow) in TPP-TR and 2D-TPP analysis. Nodes with dual color had a differential effect on thermal stability between two assays. Nodes surrounded with black circles contribute to host proteostasis as annotated by GO terms. (B,C) Heat maps showing relative protein abundances as fold change of TH6744 treatment (X-axis, 1, 3, 10 and 30 μM) compared to DMSO treatment (first column in each graph) in presence of increasing temperatures (Y-axis, 42, 45, 48, 51, 54, 57, 60, 63 and 67 °C). Half-maximal effective concentration of protein stabilization/destabilization pEC₅₀ (pEC₅₀ = −log(EC₅₀) illustrates binding affinity of TH6744 to host targets. n = 1 biological replicate. Grey squares indicate not available values. (B) Heat maps from 2D-TPP showing relative abundances of proteins typically located in the ER. (C) Heat maps from 2D-TPP showing relative abundances of proteins typically located in cytosol.