Extended Data Fig. 7: Analysis of PROTAC induced ternary complexes between the isolated protein kinase ___domain of BRAF and CRBN/DDB1 in vitro.
From: Functional characterization of a PROTAC directed against BRAF mutant V600E
a, Binding of CRBN/DDB1 to a fluorescein-labelled pomalidomide probe assessed by fluorescence polarization. b-h, Competitive displacement of fluorescein-labelled pomalidomide probe from CRBN-DDB1 (at 100 nM and 400 nM, respectively, corresponding to 80% saturated binding) by pomalidomide (Pmd) (b), P4B (c), 2 (d), 4 (e), 5 (f), 6 (g), 7 (h) assessed in the absence or in the presence of BRAF16mut(WT) kinase ___domain, or BRAF16mut(V600E) kinase ___domain. IC50 values were obtained by fitting FP signal using a variable slope - four parameter equation in GraphPad Prism. Data shown are representative of two independent experiments each performed in triplicate. Data represent mean values ± s.d.
