A tyrosine-phosphorylated 70-kDa protein binds a photoaffinity analogue of ATP and associates with both the zeta chain and CD3 components of the activated T cell antigen receptor.
An early event in T cell antigen receptor (TCR)-mediated signal transduction is the activation of a protein tyrosine kinase (PTK) pathway. An unidentified PTK activity and a kinase substrate termed ZAP-70 have previously been shown to associate with TCR zeta upon cross-linking of TCR beta. Here we report that TCR activation, by antibody cross-linking of either TCR beta or CD3 epsilon, results in the association of a PTK activity with both CD3 and TCR zeta. A number of in vitro PTK substrates are also associated with CD3 and TCR zeta, including CD3 epsilon, TCR zeta, p60fyn, p62yes, and a predominant 70-kDa protein (ZAP-70). The shared PTK activity and PTK substrates suggest that both CD3 and TCR zeta are involved in signal transduction via a shared pathway. We used [alpha-32P]gamma-azidoanilido ATP, a photoreactive analogue of ATP, to detect CD3-associated proteins that bound ATP upon TCR activation, reasoning that such proteins could represent PTKs. A 70-kDa protein bound [alpha-32P]gamma-azidoanilido ATP only upon TCR activation, and we propose that this protein and the 70-kDa PTK substrate are the same protein. Furthermore, we propose that this protein is responsible for the PTK activity observed to be associated with TCR zeta and CD3 upon TCR activation.
Wange
R L
RL
Cell Biology and Metabolism Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20892.
Kong
A N
AN
Samelson
L E
LE
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D000345:Affinity Labels; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D001386:Azides / Q000378:metabolism*; D017252:CD3 Complex; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D010449:Peptide Mapping; D010766:Phosphorylation; D010777:Photochemistry; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis / Q000378:metabolism*; D011948:Receptors, Antigen, T-Cell / Q000378:metabolism*; D015398:Signal Transduction; D013379:Substrate Specificity; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism
D000345:Affinity Labels; D000945:Antigens, Differentiation, T-Lymphocyte; D001386:Azides; D017252:CD3 Complex; D003432:Cross-Linking Reagents; D008565:Membrane Proteins; D011948:Receptors, Antigen, T-Cell; C054979:antigen T cell receptor, zeta chain; D014443:Tyrosine; C024294:ATP gamma-p-azidoanilide; D000255:Adenosine Triphosphate; D011505:Protein-Tyrosine Kinases