Analysis of galectin 1-mediated cell signaling by combined precipitation and electrophoresis techniques.
Galectin-1 (GAL1) is a beta-galactoside-binding protein that has been implicated in the regulation of viability of lymphoid cells. However, the signaling pathway governed by the binding of GAL1 to the cell membrane is not understood yet. As a first step toward the elucidation of GAL1-initiated signaling events, electrophoresis techniques such as sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and two-dimensional electrophoresis (2-DE) were used together with precipitation techniques. This allowed us to identify the membrane receptor of GAL1, and to characterize the signal resulting from the binding of GAL1 to this receptor. Our results demonstrate that the tyrosine phosphatase CD45 is the receptor for GAL1, and that the src-type tyrosine kinase Lyn is a target for the effects of GAL1/CD45 interactions in B-cells. Furthermore, these results show the usefulness of combined precipitation and electrophoresis techniques to analyze phosphotyrosine-dependent mechanisms during the study of cell functions.
Fouillit
M
M
Université Paris 13, Biochimie Cellulaire des Hémopathies Lymphoïdes, Bobigny, France.
Poirier
F
F
Monostori
E
E
Raphael
M
M
Bladier
D
D
Joubert-Caron
R
R
Caron
M
M
D015153:Blotting, Western / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005697:Galactosides / Q000378:metabolism*; D037483:Galectin 1; D006388:Hemagglutinins / Q000378:metabolism*; D006801:Humans; D037102:Lectins; D010766:Phosphorylation; D015398:Signal Transduction; D014407:Tumor Cells, Cultured; D014443:Tyrosine
D005697:Galactosides; D037483:Galectin 1; D006388:Hemagglutinins; D037102:Lectins; C007816:beta-galactoside; D014443:Tyrosine