Abstract
Recent studies have shown that, in addition to its role as an adhesion receptor, platelet endothelial cell adhesion molecule 1/CD31 becomes phosphorylated on tyrosine residues Y663 and Y686 and associates with protein tyrosine phosphatases SHP-1 and SHP-2. In this study, we screened for additional proteins which associate with phosphorylated platelet endothelial cell adhesion molecule 1, using surface plasmon resonance. We found that, besides SHP-1 and SHP-2, platelet endothelial cell adhesion molecule 1 binds the cytoplasmic signalling proteins SHIP and PLC-gamma1 via their Src homology 2 domains. Using two phosphopeptides, NSDVQpY663TEVQV and DTETVpY686SEVRK, we demonstrate differential binding of SHP-1, SHP-2, SHIP and PLC-gamma1. All four cytoplasmic signalling proteins directly associate with cellular platelet endothelial cell adhesion molecule 1, immunoprecipitated from pervanadate-stimulated THP-1 cells. These results suggest that overlapping immunoreceptor tyrosine-based inhibition motif/immunoreceptor tyrosine-based activation motif-like motifs within platelet endothelial cell adhesion molecule 1 mediate differential interactions between the Src homology 2 containing signalling proteins SHP-1, SHP-2, SHIP and PLC-gamma1.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Binding Sites
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Humans
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Intracellular Signaling Peptides and Proteins
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Isoenzymes / metabolism*
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Molecular Sequence Data
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Monocytes / metabolism
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Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases
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Phospholipase C gamma
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Phosphopeptides / metabolism
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Phosphoric Monoester Hydrolases / metabolism*
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Phosphorylation
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Phosphotyrosine / metabolism
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Platelet Endothelial Cell Adhesion Molecule-1 / chemistry
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Platelet Endothelial Cell Adhesion Molecule-1 / metabolism*
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Protein Binding
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Protein Tyrosine Phosphatase, Non-Receptor Type 11
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Protein Tyrosine Phosphatase, Non-Receptor Type 6
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Protein Tyrosine Phosphatases / metabolism*
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SH2 Domain-Containing Protein Tyrosine Phosphatases
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Sequence Homology, Amino Acid
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Signal Transduction
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Surface Plasmon Resonance
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Type C Phospholipases / metabolism*
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Vanadates / pharmacology
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src Homology Domains
Substances
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Intracellular Signaling Peptides and Proteins
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Isoenzymes
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Phosphopeptides
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Platelet Endothelial Cell Adhesion Molecule-1
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pervanadate
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Phosphotyrosine
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Vanadates
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Phosphoric Monoester Hydrolases
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PTPN11 protein, human
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PTPN6 protein, human
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Protein Tyrosine Phosphatase, Non-Receptor Type 11
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Protein Tyrosine Phosphatase, Non-Receptor Type 6
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Protein Tyrosine Phosphatases
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SH2 Domain-Containing Protein Tyrosine Phosphatases
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INPPL1 protein, human
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Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases
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Type C Phospholipases
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Phospholipase C gamma