Src family tyrosine kinase p53/56lyn, a serine kinase and Fc epsilon RI associate with alpha-galactosyl derivatives of ganglioside GD1b in rat basophilic leukemia RBL-2H3 cells.

Abstract

The monoclonal antibody (mAb) AA4 recognizes two alpha-galactosyl derivatives of the GD1b ganglioside on rat mast cells and on the rat basophilic leukemia RBL-2H3 cultured cell line. Here we demonstrate that mAb AA4 coprecipitated both protein tyrosine and serine kinases. In contrast, a monoclonal antibody to the GD3 ganglioside did not coprecipitate any kinase activity. In kinase assays of mAb AA4 immunoprecipitates there were phosphorylated proteins of 71-80, 53/56, and 41/42 kDa. All proteins were phosphorylated on tyrosine, whereas the 71-80- and 41/42-kDa proteins were also phosphorylated on serine residues. The precipitation of these proteins by mAb AA4 correlated with the presence of the alpha-galactosyl derivatives of GD1b. The 53/56-kDa proteins were identified as the Src-related tyrosine kinase p53/56lyn. The presence of p53/56lyn in the mAb AA4 immunoprecipitates was specific and was observed when several different detergents were used. The same 71-80-kDa tyrosine-phosphorylated proteins were immunoprecipitated by mAb AA4 and anti-Lyn antibodies and may play a role in the interaction of p53/56lyn with the gangliosides. Although there is a weak association of the high affinity IgE receptor with these gangliosides, the coprecipitation of p53/56lyn with mAb AA4 was not secondary to the association of this kinase with receptor. These complexes of gangliosides and several proteins that include p53/56lyn, a serine kinase, and the high affinity IgE receptor could play an important role in receptor-mediated signal transduction.

• Authors: K Minoguchi, W D Swaim, E H Berenstein, R P Siraganian
• Journal: The Journal of biological chemistry (J Biol Chem) Vol. 269 Issue 7 Pg. 5249-54 (Feb 18 1994) ISSN: 0021-9258 [Print] United States
• PMID: 8106508 (Publication Type: Journal Article)

Chemical References

• Antibodies, Monoclonal
• Galactosides
• Gangliosides
• Phosphorus Radioisotopes
• Receptors, IgE
• ganglioside, GD1b
• Protein-Tyrosine Kinases
• lyn protein-tyrosine kinase
• src-Family Kinases
• Protein Serine-Threonine Kinases

Topics

• Animals
• Antibodies, Monoclonal
• Autoradiography
• Cell Line
• Electrophoresis, Polyacrylamide Gel
• Galactosides (analysis, metabolism)
• Gangliosides (isolation & purification, metabolism)
• Immunoblotting
• Leukemia, Basophilic, Acute
• Mast Cells (enzymology)
• Molecular Weight
• Phosphorus Radioisotopes
• Protein Serine-Threonine Kinases (isolation & purification, metabolism)
• Protein-Tyrosine Kinases (isolation & purification, metabolism)
• Rats
• Receptors, IgE (isolation & purification, metabolism)
• Tumor Cells, Cultured
• src-Family Kinases