Phosphorylation of two cytosolic proteins. An early event of T-cell activation.
T lymphocytes can be activated to proliferate by triggering the T-cell antigen-receptor complex (CD3-Ti) with anti-CD3 (Cluster of Differentiation 3) monoclonal antibody (mAb) or with the mitogenic lectin phytohaemagglutinin A (PHA). We have investigated the relationship between lymphocyte activation and protein phosphorylation in the human leukaemic T-cell line Jurkat. Incubation of 32P-labelled Jurkat cells with anti-CD3 mAb or PHA induced the phosphorylation of two cytosolic proteins that migrate with apparent Mr values of 21,000 (pp21) and 23,000 (pp23) and pI values of 5.1 and 5.0 respectively. Peptide mapping of the two proteins produced the same phosphopeptides pattern, suggesting that pp21 and pp23 are closely related. The phosphorylation of pp21 and pp23 induced by anti-CD3 mAb appeared to be transient, since it was already detected 2 min after the addition of the mAb, reached a maximum at 10 min and recovered its basal level after 1 h. Phosphorylation of pp21 and pp23 could also be elicited by the Ca2+ ionophore A23187 and sodium orthovanadate (Na3VO4), two agents that bypass the T-cell-receptor complex and produced an increase in cytosolic Ca2+ concentration. In addition, we found that vanadate, like the Ca2+ ionophore, induced the secretion of interleukin-2 (IL-2) when used in combination with a submitogenic concentration of the phorbol ester 12-O-tetradecanoylphorbol 13-acetate. These results show that the Ca2+-dependent phosphorylation of pp21 and pp23 represents an early event in the process of signal transduction through the CD3-Ti receptor complex.
Peyron
J F
JF
INSERM U 210, Faculté de Médecine (Pasteur), Nice, France.
Aussel
C
C
Ferrua
B
B
Häring
H
H
Fehlmann
M
M
D000001:Calcimycin / Q000494:pharmacology; D002460:Cell Line; D003600:Cytosol / Q000378:metabolism*; D006801:Humans; D007376:Interleukin-2 / Q000494:pharmacology; D008213:Lymphocyte Activation; D010449:Peptide Mapping; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*; D013601:T-Lymphocytes / Q000502:physiology*; D014407:Tumor Cells, Cultured / Q000187:drug effects; D014638:Vanadates / Q000494:pharmacology
D007376:Interleukin-2; D011506:Proteins; D000001:Calcimycin; D014638:Vanadates