Fyn Kinase-directed Activation of SH2 Domain-containing Protein-tyrosine Phosphatase SHP-2 by Gi Protein-coupled Receptors in Madin-Darby Canine Kidney Cells

• Published in: The Journal of biological chemistry Vol. 274; no. 18; pp. 12401 - 12407
• Main Authors: Tang, H, Zhao, Z J, Huang, X Y, Landon, E J, Inagami, T
• Format: Journal Article
• Language: English
• Published: United States American Society for Biochemistry and Molecular Biology 30-04-1999
• Subjects: Adaptor Proteins, Signal Transducing; Amino Acid Sequence; Animals; Cell Line; Dogs; Enzyme Activation; GRB2 Adaptor Protein; GTP-Binding Protein alpha Subunits, Gi-Go - metabolism; Intracellular Signaling Peptides and Proteins; Molecular Sequence Data; Pertussis Toxin; Protein Binding; Protein Tyrosine Phosphatase, Non-Receptor Type 11; Protein Tyrosine Phosphatase, Non-Receptor Type 6; Protein Tyrosine Phosphatases - metabolism; Protein-Tyrosine Kinases - metabolism; Proteins - metabolism; Receptors, Adrenergic, alpha-2 - metabolism; Receptors, Cell Surface - metabolism; Receptors, G-Protein-Coupled; Receptors, Lysophosphatidic Acid; SH2 Domain-Containing Protein Tyrosine Phosphatases; src Homology Domains; src-Family Kinases - metabolism; Virulence Factors, Bordetella - pharmacology
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.274.18.12401

SHP-2, an SH2 domain-containing protein-tyrosine phosphatase, plays an important role in receptor tyrosine kinase-regulated
cell proliferation and differentiation. Little is known about the activation mechanisms and the participation of SHP-2 in
the activity of G protein-coupled receptors lacking intrinsic tyrosine kinase activity. We show that the activity of SHP-2
(but not SHP-1) is specifically stimulated by the selective α 2A -adrenergic receptor agonist UK14304 and by lysophosphatidic acid (LPA) in Madin-Darby canine kidney (MDCK) cells. UK14304
and LPA promote the tyrosine phosphorylation of SHP-2 and its association with Grb2. The agonist-induced direct interaction
of Grb2 with SHP-2 is mediated by the SH2 domain of Grb2 and the tyrosine phosphorylation of SHP-2. Rapid activation of Src
family kinase by UK14304 preceded the SHP-2 activation. Among the Src family members (Src, Fyn, Lck, Yes, and Lyn) present
in MDCK cells, Fyn was the only one specifically associated with SHP-2, and the physical interaction between them, which requires
the Src family kinase activity, was increased in response to the agonists. Pertussis toxin, PP1 (a selective Src family kinase
inhibitor), or overexpression of a catalytically inactive mutant of Fyn blocked the UK14304- or LPA-stimulated activity of
SHP-2, SHP-2 tyrosine phosphorylation, and SHP-2 association with Grb2. Therefore, we have demonstrated for the first time
that the activation of SHP-2 by these G i protein-coupled receptors requires Fyn kinase and that there is a specific physical interaction of Fyn kinase with SHP-2 in
MDCK cells.