Fig. 7
From: Fructose-1,6-bisphosphate couples glycolytic flux to activation of Ras
Fru1,6bisP binds to Sos1 and disrupts the Sos1/H-Ras complex. a, Biolayer interferometry (BLI) measurements showing the disruption of the Sos1/H-Ras complex. His-tagged H-Ras was coupled to Ni2+-coated biosensors and loaded with 0.5 µM of non-tagged Sos1 (association phase). Subsequently, dissociation of Sos1 was monitored in buffer containing increasing concentrations of Fru1,6bisP (0, 2, 5, 10, 20, 50, and 100 mM as labeled on the curves). The rates of Sos1 dissociation are dependent on the Fru1,6bisP concentration. b Titration curve showing the influence of Fru1,6bisP concentration on Sos1/H-Ras complex dissociation. Plotting the BLI signal amplitude as shown in panel a after 150 s of complex dissociation (R; mean ± s.d.; n = 3 independent experiments) vs. the Fru1,6bisP concentration yields a binding curve that was fitted on a Langmuir binding model to obtain an apparent K D (K Dapp) value (±s.e.). c Titration curves for binding of Fru1,6bisP to either Sos1 or H-Ras. Melting temperatures (T m) of Sos1 and H-Ras at different ligand concentrations were obtained via thermal shift assays (n = 3 independent experiments). Plotting T m values (mean ± s.d.) vs. ligand concentration yielded binding curves that were fitted on a Langmuir binding model to obtain K D values (±s.e.)
