期刊
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
卷 107, 期 51, 页码 22128-22133出版社
NATL ACAD SCIENCES
DOI: 10.1073/pnas.1016388108
关键词
Forster's resonant energy transfer; receptor trafficking; Lck kinase
资金
- Jane Coffin Childs Memorial Fund for Medical Research
Phosphorylation of the T-cell receptor complex (TcR/CD3) mediates the survival and antigen-induced activation of T cells. TcR/CD3 phosphorylation is usually monitored using phospho-specific antibodies, which precludes dynamic measurements. Here, we have developed genetically encoded, live-cell reporters that enable simultaneous monitoring of the phosphorylation state and intracellular trafficking of CD3 zeta, the major signal-transducing subunit of the TcR/CD3. We show that these reporters provide accurate read-outs of TcR/CD3 phosphorylation and are sensitive to the local balance of kinase and phosphatase activities acting upon TcR/CD3. Using these reporters, we demonstrate that, in addition to the expected activation-dependent phosphorylation at the plasma membrane, tyrosine-phosphorylated CD3 zeta accumulates on endosomal vesicles distinct from lysosomes. These results suggest that an intracellular pool of phosphorylated CD3 zeta may help to sustain TcR/CD3 signaling after the receptor internalization.
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