1. Two clusters of residues at the docking groove of mitogen-activated protein kinases differentially mediate their functional interaction with the tyrosine phosphatases PTP-SL and STEP
- Author
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Carmen Blanco-Aparicio, Juan José Muñoz, Céline Tárrega, and Rafael Pulido
- Subjects
Time Factors ,Protein tyrosine phosphatase ,environment and public health ,Biochemistry ,p38 Mitogen-Activated Protein Kinases ,Mice ,Cytosol ,Tyrosine ,Phosphorylation ,Cells, Cultured ,Glutathione Transferase ,Mitogen-Activated Protein Kinase 1 ,Kinase ,Intracellular Signaling Peptides and Proteins ,Protein Tyrosine Phosphatases, Non-Receptor ,Receptor-Like Protein Tyrosine Phosphatases ,Drosophila ,Electrophoresis, Polyacrylamide Gel ,Signal transduction ,Mitogen-Activated Protein Kinases ,hormones, hormone substitutes, and hormone antagonists ,Protein Binding ,Signal Transduction ,animal structures ,MAP Kinase Signaling System ,Recombinant Fusion Proteins ,Phosphatase ,Immunoblotting ,Molecular Sequence Data ,Nerve Tissue Proteins ,Saccharomyces cerevisiae ,Biology ,Transfection ,Cell Line ,Animals ,Humans ,Amino Acid Sequence ,Receptor-Like Protein Tyrosine Phosphatases, Class 7 ,Molecular Biology ,Binding Sites ,Dose-Response Relationship, Drug ,Epidermal Growth Factor ,Sequence Homology, Amino Acid ,Cell Biology ,Precipitin Tests ,Protein Structure, Tertiary ,enzymes and coenzymes (carbohydrates) ,Microscopy, Fluorescence ,Docking (molecular) ,Mutation ,Mutagenesis, Site-Directed ,Protein Tyrosine Phosphatases - Abstract
Regulated function of mitogen-activated protein (MAP) kinases involves their selective association through docking sites with both activating MAP kinase kinases and inactivating phosphatases, including dual specificity and protein-tyrosine phosphatases (PTP). Site-directed mutagenesis on the mammalian MAP kinases ERK2 and p38alpha identified within their C-terminal docking grooves two clusters of residues important for association with their regulatory PTPs, PTP-SL and STEP. ERK2 and p38alpha mutations that resembled the sevenmaker gain-of-function mutation in the Rolled D. melanogaster ERK2 homologue failed to associate with PTP-SL, were not retained in the cytosol, and were poorly inactivated by this PTP. Additional ERK2 mutations at the docking groove showed deficient association and dephosphorylation by PTP-SL, although their cytosolic retention was unaffected. Other ERK2 mutations, resembling gain-of-function mutations in the FUS3 yeast ERK2 homologue, associated to PTP-SL and were inactivated normally by this PTP. Our results demonstrate that mutations at distinct regions of the docking groove of ERK2 and p38alpha differentially affect their association and regulation by the PTP-SL and STEP PTPs.
- Published
- 2001