Fig. 7: Binding and affinity measurements of abemaciclib towards HIPK2, HIPK3, DYRK1A, and Cdk4/CycD3.
From: Abemaciclib is a potent inhibitor of DYRK1A and HIP kinases involved in transcriptional regulation
a Surface plasmon resonance (SPR) measurements were performed using single-cycle kinetics. Kinases were immobilized to the chip surface by amine coupling. Abemaciclib was injected at increasing concentrations of 0.55, 1.2, 2.6, 5.8, 12.8, 28.2, 62, 136.4, and 300 nM for 120 s, followed by dissociation for 900 s. Dissociation constants were determined based on fits applying a 1:1 interaction model. b Thermal protein stability measurements were performed using the NanoDSF technique. Proteins were diluted to 5 µM concentration in kinase buffer and incubated with 1, 10, or 100 µM abemaciclib for 10 min. Measurements were performed in duplicates (n = 2 biologically independent samples) and are depicted as mean. c Thermal protein stability measurements were performed as in (b) using ADP·Mg2+, ATP·Mg2+, or abemaciclib. ∆Tm (max.–min. in °C) is depicted as mean. For raw data see Supplementary Table 3. Source data are provided as a Source Data file.
