title: Differential association of cytoplasmic signalling molecules SHP-1, SHP-2, SHIP and phospholipase C-gamma1 with PECAM-1/CD31.
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.
authors.affiliation: Division of Immunity and Infection, University of Birmingham, UK.
authors.forename: N J
authors.initials: NJ
authors.lastname: Pumphrey
authors.forename: V
authors.initials: V
authors.lastname: Taylor
authors.forename: S
authors.initials: S
authors.lastname: Freeman
authors.forename: M R
authors.initials: MR
authors.lastname: Douglas
authors.forename: P F
authors.initials: PF
authors.lastname: Bradfield
authors.forename: S P
authors.initials: SP
authors.lastname: Young
authors.forename: J M
authors.initials: JM
authors.lastname: Lord
authors.forename: M J
authors.initials: MJ
authors.lastname: Wakelam
authors.forename: I N
authors.initials: IN
authors.lastname: Bird
authors.forename: M
authors.initials: M
authors.lastname: Salmon
authors.forename: C D
authors.initials: CD
authors.lastname: Buckley
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes / Q000378:metabolism*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000378:metabolism; D000072183:Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases; D051966:Phospholipase C gamma; D010748:Phosphopeptides / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases / Q000378:metabolism*; D054593:SH2 Domain-Containing Protein Tyrosine Phosphatases; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction; D020349:Surface Plasmon Resonance; D010738:Type C Phospholipases / Q000378:metabolism*; D014638:Vanadates / Q000494:pharmacology; D018909:src Homology Domains
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D007527:Isoenzymes; D010748:Phosphopeptides; D019408:Platelet Endothelial Cell Adhesion Molecule-1; C076715:pervanadate; D019000:Phosphotyrosine; D014638:Vanadates; D010744:Phosphoric Monoester Hydrolases; C516724:PTPN11 protein, human; C509249:PTPN6 protein, human; D054592:Protein Tyrosine Phosphatase, Non-Receptor Type 11; D053824:Protein Tyrosine Phosphatase, Non-Receptor Type 6; D017027:Protein Tyrosine Phosphatases; D054593:SH2 Domain-Containing Protein Tyrosine Phosphatases; C098706:INPPL1 protein, human; D000072183:Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases; D010738:Type C Phospholipases; D051966:Phospholipase C gamma