Fig. 3: G12D inhibitors bind to both GDP-bound and GMPPNP-bound KRAS.
From: KRAS(G12D) can be targeted by potent inhibitors via formation of salt bridge
a Upper panel: Fo-Fc map of TH-Z827, Asp12, and GDP (PDB ID: 7EWA, monomer A). Fo-Fc map of TH-Z827, Asp12, and the GTP analog GMPPNP (PDB ID: 7EWA, monomer B). Lower panel: Fo-Fc map of TH-Z835, Asp12, and GDP (PDB ID: 7EWB, monomer A). Fo-Fc map of TH-Z835, Asp12, and GMPPNP (PDB ID: 7EWB, monomer B). In all graphs, the 1.5 σ Fo-Fc maps of all shown elements are shown in gray mesh. The color scheme of 2.5 σ Fo-Fc maps is: blue for inhibitors, pink for Asp12, and yellow for nucleotides. b KRAS(G12D) conformational change analysis for drug-free GDP-bound protein (PDB: 4EPR), drug-free GMPPNP-bound protein (PDB: 5USJ), TH-Z835 and GDP-bound protein (PDB: 7EWB, chain A), and TH-Z835 and GMPPNP-bound protein (PDB: 7EWB, chain B). c EDTA-mediated competition between fluorescently labeled mantGDP loaded on KRAS and free nucleotide (GDP or GTP). The experiment was carried out with KRAS(G12D) alone or with KRAS(G12D) treated with TH-Z835. d Inhibitory activity of TH-Z835 measured by SOS-catalyzed nucleotide exchange assays with GMPPNP as the incoming nucleotide.
