Figure EV2. hGBP1_F polymerization is dependent on GTP, and subsequent GDP hydrolysis and its dynamics are modified by the C‐terminal 3R stretch.

• A
  Polymerization of 10 μM hGBP1_F ^R584‐586A was monitored over time by absorption spectroscopy at 350 nm after admixture of 2 mM (GTP, GppNHp, GTPγS) or 250 μM (GDP·AlF_X) nucleotide.
• B
  Polymerization of different hGBP1 variants (at 10 μM) induced by 2 mM GTP was monitored over time by absorption spectroscopy at 350 nm. Absorbance signals were superimposed with nucleotide composition analyzed at defined time points of the corresponding sample. Blots for hGBP1_F and hGBP1_F ^R584‐586A are the same as shown in Fig 3A.
• C
  GTP turnover numbers were determined for different hGBP1 variants. Turnover numbers for both the slow and the fast phases of GTP hydrolysis (1st phase [1.], 2nd phase [2.]) were quantified during hGBP1_F and hGBP1_F ^R584‐586A polymerization. Combined data from three independent experiments are shown as mean turnover numbers ± SEM. Significance was determined by two‐way ANOVA with Tukey's multiple comparison test. n.s., not significant; **P ≤ 0.01; ****P ≤ 0.0001.

Source data are available online for this figure.