Fig 1: Specific targeting of DHS.Active site with bound a spermidine (SPD) b N1-guanyl-1,7-diaminoheptane (GC7), and c VT00065 covalently linked to K329. d Proposed simplified mechanism of reaction leading to linking of VT00065 to the side chain of lysine. e In vitro deoxyhypusination assay with Western blot readout demonstrating weak but dose-dependent inhibitory effect of VT00065. f VT00155 fragment (pyroquilone), shown as sticks, is bound to the DHS interface with two molecules in close proximity, suggesting the possibility for linking. g DMP7 molecule bound at the DHS tetramer interface is shown as purple sticks. h The close-up view on the DMP7 and its binding pocket with residues involved in binding shown as sticks. i Electron density map shown as blue mesh contoured at 1σ level. j Western blot-based DHS activity assay showing no impact of DMP7 on the catalytic activity of DHS. k The same assay as in (j), showing stabilizing effect of DMP7 after prolonged incubation of DHS in the assay conditions (control samples were also incubates at RT for 3 h before analysis). For e, j, k: for deoxyhypusination detection, monoclonal rabbit FabHpu98 (#PABL-582) antibody was used (Creative Biolabs). Data are mean ± SEM of biological triplicates where “*” indicates a statistically difference from the control sample with P < 0.005 determined by one-way ANOVA with post hoc Dunnett’s tests. Below graphs, representative Western blots and loading controls are shown. All of the replicates with statistical analysis are presented in Supplementary Figs. 3 and 4.