• Register
X
Forgot Password

If you have forgotten your password you can enter your email here and get a temporary password sent to your email.

X

Leaving Community

Are you sure you want to leave this community? Leaving the community will revoke any permissions you have been granted in this community.

No
Yes

Characterization of ERK1 activation site mutants and the effect on recognition by MEK1 and MEK2.

To discern MEK1 and MEK2 specificity for their substrate, extracellular signal-regulated kinase (ERK), site-directed mutagenesis was performed on the amino acid residues flanking the regulatory phosphorylation sites of ERK1. These ERK1 mutants were analyzed for the ability to act as a substrate for MEK1 and MEK2. Based on both phosphorylation and activation analyses, the mutants could be divided into four classes: 1) dramatically decreased phosphorylation and activation, 2) enhanced basal kinase activity, 3) preferentially enhanced phosphorylation of tyrosine and decreased phosphorylation of threonine, and 4) increased threonine phosphorylation with an increase in activation. In general, the residues proximal to the regulatory phosphorylation sites of ERK1 had greater influence on both phosphorylation and activation. This is consistent with the highly specific recognition of the ERK1 regulatory sites by MEK. Mutation of Arg-208 or Thr-207 to an alanine residue significantly altered the relative phosphorylation on Thr-202 and Tyr-204. The Arg-208 to alanine mutant increased the phosphorylation of Tyr-204 approximately 4-fold yet almost completely eliminated the phosphorylation on Thr-202. In contrast, mutation of Gly-199 to alanine resulted in an increased phosphorylation of Thr-202 relative to Tyr-204. This suggests that both Gly-199 and Arg-208 play important roles in determining the relative phosphorylation of Thr-202 and Tyr-204. Our results demonstrate that residues in the phosphorylation lip of ERK play an important role in the recognition and phosphorylation by MEK.

Pubmed ID: 8626767

Authors

  • Butch ER
  • Guan KL

Journal

The Journal of biological chemistry

Publication Data

February 23, 1996

Associated Grants

  • Agency: NIA NIH HHS, Id: 5T32AG00114
  • Agency: NIGMS NIH HHS, Id: GM51586

Mesh Terms

  • Amino Acid Sequence
  • Binding Sites
  • Calcium-Calmodulin-Dependent Protein Kinases
  • Cloning, Molecular
  • Conserved Sequence
  • Enzyme Activation
  • Humans
  • MAP Kinase Kinase 1
  • MAP Kinase Kinase 2
  • Mitogen-Activated Protein Kinase 3
  • Mitogen-Activated Protein Kinase Kinases
  • Mitogen-Activated Protein Kinases
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Phosphorylation
  • Protein-Serine-Threonine Kinases
  • Protein-Tyrosine Kinases
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • Regulatory Sequences, Nucleic Acid
  • Restriction Mapping
  • Tyrosine