Interaction of Mitogen-activated Protein Kinases with the Kinase Interaction Motif of the Tyrosine Phosphatase PTP-SL Provides Substrate Specificity and Retains ERK2 in the Cytoplasm

6533b838fe1ef96bd12a3ef6

RESEARCH PRODUCT

Interaction of Mitogen-activated Protein Kinases with the Kinase Interaction Motif of the Tyrosine Phosphatase PTP-SL Provides Substrate Specificity and Retains ERK2 in the Cytoplasm

Rafael Pulido ÁNgel Zúñiga Josefa ÚBeda Josema Torres

subject

Cytoplasm animal structures Protein Kinase C-alpha Recombinant Fusion Proteins Cèl·lules Nerve Tissue Proteins Protein tyrosine phosphatase Mitogen-activated protein kinase kinase Transfection environment and public health Biochemistry SH3 domain Receptor tyrosine kinase MAP2K7 Substrate Specificity Serine Animals c-Raf Amino Acid Sequence Molecular Biology Protein Kinase C Sequence Deletion Mitogen-Activated Protein Kinase 1 Binding Sites Mitogen-Activated Protein Kinase 3 biology Cyclin-dependent kinase 2 Intracellular Signaling Peptides and Proteins JNK Mitogen-Activated Protein Kinases Cell Biology Cyclic AMP-Dependent Protein Kinases Isoenzymes enzymes and coenzymes (carbohydrates)Kinetics Biochemistry Amino Acid Substitution COS Cells Calcium-Calmodulin-Dependent Protein Kinases biology.protein Mutagenesis Site-Directed Cyclin-dependent kinase 9 Cattle Mitogen-Activated Protein Kinases Protein Tyrosine Phosphatases Proteïnes

description

ERK1 and ERK2 associate with the tyrosine phosphatase PTP-SL through a kinase interaction motif (KIM) located in the juxtamembrane region of PTP-SL. A glutathione S-transferase (GST)-PTP-SL fusion protein containing the KIM associated with ERK1 and ERK2 as well as with p38/HOG, but not with the related JNK1 kinase or with protein kinase A or C. Accordingly, ERK2 showed in vitro substrate specificity to phosphorylate GST-PTP-SL in comparison with GST-c-Jun. Furthermore, tyrosine dephosphorylation of ERK2 by the PTP-SLDeltaKIM mutant was impaired. The in vitro association of ERK1/2 with GST-PTP-SL was highly stable; however, low concentrations of nucleotides partially dissociated the ERK1/2.PTP-SL complex. Partial deletions of the KIM abrogated the association of PTP-SL with ERK1/2, indicating that KIM integrity is required for interaction. Amino acid substitution analysis revealed that Arg and Leu residues within the KIM are essential for the interaction and suggested a regulatory role for Ser(231). Finally, coexpression of PTP-SL and ERK2 in COS-7 cells resulted in the retention of ERK2 in the cytoplasm in a KIM-dependent manner. Our results demonstrate that the noncatalytic region of PTP-SL associates with mitogen-activated protein kinases with high affinity and specificity, providing a mechanism for substrate specificity, and suggest a role for PTP-SL in the regulation of mitogen-activated protein kinase translocation to the nucleus upon activation.

year	journal	country	edition	language
1999-01-01

10.1074/jbc.274.31.21900 https://hdl.handle.net/10550/84237