Identification of Tek/Tie2 binding partners. Binding to a multifunctional docking site mediates cell survival and migration

N Jones; Z Master; J Jones; D Bouchard; Y Gunji; H Sasaki; R Daly; K Alitalo; D J Dumont

doi:10.1074/jbc.274.43.30896

Identification of Tek/Tie2 binding partners. Binding to a multifunctional docking site mediates cell survival and migration

J Biol Chem. 1999 Oct 22;274(43):30896-905. doi: 10.1074/jbc.274.43.30896.

Authors

N Jones¹, Z Master, J Jones, D Bouchard, Y Gunji, H Sasaki, R Daly, K Alitalo, D J Dumont

Affiliation

¹ Division of Cancer Biology Research, Sunnybrook and Women's College Health Sciences Centre, University of Toronto, Toronto, Ontario M4N 3M5.

PMID: 10521483
DOI: 10.1074/jbc.274.43.30896

Abstract

The Tek/Tie2 receptor tyrosine kinase plays a pivotal role in vascular and hematopoietic development. To study the signal transduction pathways that are mediated by this receptor, we have used the yeast two-hybrid system to identify signaling molecules that associate with the phosphorylated Tek receptor. Using this approach, we demonstrate that five molecules, Grb2, Grb7, Grb14, Shp2, and the p85 subunit of phosphatidylinositol 3-kinase can interact with Tek in a phosphotyrosine-dependent manner through their SH2 domains. Mapping of the binding sites of these molecules on Tek reveals the presence of a multisubstrate docking site in the carboxyl tail of Tek (Tyr(1100)). Mutation of this site abrogates binding of Grb2 and Grb7 to Tek in vivo, and this site is required for tyrosine phosphorylation of Grb7 and p85 in vivo. Furthermore, stimulation of Tek-expressing cells with Angiopoietin-1 results in phosphorylation of both Tek and p85 and in activation of endothelial cell migration and survival pathways that are dependent in part on phosphatidylinositol 3-kinase. Taken together, these results demonstrate that Angiopoietin-1-induced signaling from the Tek receptor is mediated by a multifunctional docking site that is responsible for activation of both cell migration and cell survival pathways.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adaptor Proteins, Signal Transducing*
Amino Acid Sequence
Animals
Binding Sites
Cell Movement / physiology
Cell Survival / physiology
Embryo, Mammalian
ErbB Receptors / metabolism*
GRB2 Adaptor Protein
GRB7 Adaptor Protein
Gene Library
Intracellular Signaling Peptides and Proteins
Lung / metabolism
Mice
Molecular Sequence Data
Myocardium / metabolism
Phosphatidylinositol 3-Kinases / chemistry
Phosphatidylinositol 3-Kinases / metabolism
Protein Binding
Protein Tyrosine Phosphatase, Non-Receptor Type 11
Protein Tyrosine Phosphatase, Non-Receptor Type 6
Protein Tyrosine Phosphatases / metabolism
Proteins / metabolism
Receptor Protein-Tyrosine Kinases / metabolism*
Receptor, TIE-2
Recombinant Fusion Proteins / metabolism
SH2 Domain-Containing Protein Tyrosine Phosphatases
Saccharomyces cerevisiae / genetics
Saccharomyces cerevisiae / growth & development
src Homology Domains

Substances

Adaptor Proteins, Signal Transducing
GRB2 Adaptor Protein
Grb14 protein, mouse
Grb2 protein, mouse
Grb7 protein, mouse
Intracellular Signaling Peptides and Proteins
Proteins
Recombinant Fusion Proteins
GRB7 Adaptor Protein
ErbB Receptors
Receptor Protein-Tyrosine Kinases
Receptor, TIE-2
Protein Tyrosine Phosphatase, Non-Receptor Type 11
Protein Tyrosine Phosphatase, Non-Receptor Type 6
Protein Tyrosine Phosphatases
Ptpn11 protein, mouse
Ptpn6 protein, mouse
SH2 Domain-Containing Protein Tyrosine Phosphatases