Fig. 3
From: Modulating multi-functional ERK complexes by covalent targeting of a recruitment site in vivo
BI-78D3 exhibits a unique selectivity towards ERK1/2. a Kinetic behavior of ERK2 inactivation by BI-78D3. Data were fit to Eq. (1) as described in the methods section and according to the two-step mechanism of activation shown in Scheme 1 (data represents one experiment out of two repetitions; both produced consistent kinetic parameters, values of Ki and kinact ± standard error). b The final frame of a 100 ns molecular dynamics trajectory for BI-78D3 binding to ERK2 (PDB 4ERK). The figure was generated using UCSF Chimera software (https://www.cgl.ucsf.edu/chimera/)70. c Sequence alignment of Human MAPKs encompassing the D-recruitment site. The cysteines corresponding to C159 in ERK2 (targeted by BI-78D3) are indicated by the black rectangle; the numbering corresponds to human ERK1 sequence. C178 in Homo sapiens ERK1 corresponds to C161 in Homo sapiens ERK2 and C159 in Rattus norvegicus ERK2. d Reversibility of JNK1, but not ERK2 inhibition by BI-78D3. Each enzyme (5 µM) was treated with BI-78D3 (100 µM) or DMSO (control) for 1 h. The activity of each enzyme was estimated before and after excessive dialysis (data are from three independent experiments, and bars represent mean ± SD)
