A single amino-acid change in ERK1/2 makes the enzyme susceptible to PP1 derivatives
S Endo, Y Satoh, K Shah, K Takishima - Biochemical and biophysical …, 2006 - Elsevier
S Endo, Y Satoh, K Shah, K Takishima
Biochemical and biophysical research communications, 2006•ElsevierWe generated extracellular signal-regulated kinase 1/2 (ERK1/2) mutants by introducing a
single amino-acid substitution in subdomain V of the catalytic domain and then examined
the susceptibility of these mutants to PP1 derivatives originally designed as Src inhibitors.
Substituting smaller amino acids (alanine [Ala (A)] or glycine [Gly (G)]) for glutamine [Gln (Q)]
in subdomain V drastically increased the susceptibility of ERK1/2 to 1-naphthyl PP1 (1NA-
PP1). Wild-type ERK1/2 was resistant to 1NA-PP1 inhibition. ERK1 (Q122A) and ERK2 …
single amino-acid substitution in subdomain V of the catalytic domain and then examined
the susceptibility of these mutants to PP1 derivatives originally designed as Src inhibitors.
Substituting smaller amino acids (alanine [Ala (A)] or glycine [Gly (G)]) for glutamine [Gln (Q)]
in subdomain V drastically increased the susceptibility of ERK1/2 to 1-naphthyl PP1 (1NA-
PP1). Wild-type ERK1/2 was resistant to 1NA-PP1 inhibition. ERK1 (Q122A) and ERK2 …
We generated extracellular signal-regulated kinase 1/2 (ERK1/2) mutants by introducing a single amino-acid substitution in subdomain V of the catalytic domain and then examined the susceptibility of these mutants to PP1 derivatives originally designed as Src inhibitors. Substituting smaller amino acids (alanine [Ala (A)] or glycine [Gly (G)]) for glutamine [Gln (Q)] in subdomain V drastically increased the susceptibility of ERK1/2 to 1-naphthyl PP1 (1NA-PP1). Wild-type ERK1/2 was resistant to 1NA-PP1 inhibition. ERK1(Q122A) and ERK2(Q103A) were inhibited by 1NA-PP1 at IC50 values of 1.7±0.13 and 2.1±0.18μM, respectively. ERK1(Q122G) and ERK2(Q103G) were inhibited by 1NA-PP1 with IC50 values of 3.6±0.26 and 18±2.2μM, respectively. Other derivatives of PP1 (1-naphthylmethyl PP1 and 2-naphthylmethyl PP1) did not significantly inhibit ERK1/2 and its various mutants. In addition, these ERK1/2 mutants were activated by TPA when they were expressed in mammalian cells. These results suggest that the Gln residue of subdomain V is important in determining the susceptibility of ERK1/2 to 1NA-PP1 without significant changes in their enzymatic characteristics.
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