Figure 5. Ca2+ binding and myristoylation independently affect the interaction between CHP3 and MBP-CBD as measured with microscale thermophoresis (MST).
The interaction of CHP3 (A, B) and mCHP3 (C, D) with MBP-CBD was measured with three biological replicates shown in different colours (each with five technical replicates) in the presence of either Ca2+ (A, C) or Mg2+ (B, D). The combined data from individual experiments were fitted with a one-site binding model using global non-linear regression. (E) 95% confidence intervals (Δχ2 of 3.84) of KD’s for CHP3:CBD and mCHP3:CBD calculated with profile likelihood method. Raw MST traces are shown in Figure 5—figure supplement 1.
Figure 5—figure supplement 1. Normalized microscale thermophoresis (MST) fluorescence timetraces of CHP3 and mCHP3 with MBP-CBD in the presence of either Mg2+ or Ca2+.
Each graph contains measurements of one biological replicate (indicated with label (1), (2), (3)) with five technical replicates.