scholarly journals ZAP-70 Association with T Cell Receptor ζ (TCRζ): Fluorescence Imaging of Dynamic Changes upon Cellular Stimulation

1998 ◽  
Vol 143 (3) ◽  
pp. 613-624 ◽  
Author(s):  
Joanne Sloan-Lancaster ◽  
John Presley ◽  
Jan Ellenberg ◽  
Tetsuo Yamazaki ◽  
Jennifer Lippincott-Schwartz ◽  
...  

The nonreceptor protein tyrosine kinase ZAP-70 is a critical enzyme required for successful T lymphocyte activation. After antigenic stimulation, ZAP-70 rapidly associates with T cell receptor (TCR) subunits. The kinetics of its translocation to the cell surface, the properties of its specific interaction with the TCRζ chain expressed as a chimeric protein (TTζ and Tζζ), and its mobility in different intracellular compartments were studied in individual live HeLa cells, using ZAP-70 and Tζζ fused to green fluorescent protein (ZAP-70 GFP and Tζζ–GFP, respectively). Time-lapse imaging using confocal microscopy indicated that the activation-induced redistribution of ZAP-70 to the plasma membrane, after a delayed onset, is of long duration. The presence of the TCRζ chain is critical for the redistribution, which is enhanced when an active form of the protein tyrosine kinase Lck is coexpressed. Binding specificity to TTζ was indicated using mutant ZAP-70 GFPs and a truncated ζ chimera. Photobleaching techniques revealed that ZAP-70 GFP has decreased mobility at the plasma membrane, in contrast to its rapid mobility in the cytosol and nucleus. Tζζ– GFP is relatively immobile, while peripherally located ZAP-70 in stimulated cells is less mobile than cytosolic ZAP-70 in unstimulated cells, a phenotype confirmed by determining the respective diffusion constants. Examination of the specific molecular association of signaling proteins using these approaches has provided new insights into the TCRζ–ZAP-70 interaction and will be a powerful tool for continuing studies of lymphocyte activation.

Nature ◽  
2007 ◽  
Vol 446 (7137) ◽  
pp. 824-824
Author(s):  
Marcos H. Hatada ◽  
Xiaode Lu ◽  
Ellen R. Laird ◽  
Jeremy Green ◽  
Jay P. Morgenstern ◽  
...  

Cell ◽  
1991 ◽  
Vol 65 (2) ◽  
pp. 281-291 ◽  
Author(s):  
Michael P. Cooke ◽  
Kristin M. Abraham ◽  
Katherine A. Forbush ◽  
Roger M. Perimutter

Nature ◽  
1995 ◽  
Vol 377 (6544) ◽  
pp. 32-38 ◽  
Author(s):  
Marcos H. Hatada ◽  
Xiaode Lu ◽  
Ellen R. Laird ◽  
Jeremy Green ◽  
Jay P. Morgenstern ◽  
...  

Nature ◽  
1993 ◽  
Vol 365 (6446) ◽  
pp. 552-554 ◽  
Author(s):  
Steven J. Anderson ◽  
Steven D. Levin ◽  
Roger M. Perlmutter

1996 ◽  
Vol 184 (2) ◽  
pp. 365-376 ◽  
Author(s):  
V A Boussiotis ◽  
D L Barber ◽  
B J Lee ◽  
J G Gribben ◽  
G J Freeman ◽  
...  

When stimulated through their antigen receptor, without costimulation, T cells enter a state of antigen-specific unresponsiveness, termed anergy. B7-mediated costimulation, signaling via CD28, is sufficient to prevent the induction of anergy. Here we show that ligation of T cell receptor (TCR) by alloantigen alone, which results in anergy, activates tyrosine phosphorylation of TCR zeta and its association with fyn. In contrast, TCR ligation in the presence of B7 costimulation, which results in productive immunity, activates tyrosine phosphorylation of TCR zeta and CD3 chains, which associate with activated lck and zeta-associated protein (ZAP) 70. Under these conditions, CD28 associates with activated lck and TCR zeta. These data suggest that the induction of anergy is an active signaling process characterized by the association of TCR zeta and fyn. In addition, CD28-mediated costimulation may prevent the induction of anergy by facilitating the effective association of TCR zeta and CD3 epsilon with the critical protein tyrosine kinase lck, and the subsequent recruitment of ZAP-70. Strategies to inhibit or activate TCR-associated, specific protein tyrosine kinase-mediated pathways may provide a basis for drug development with potential applications in the fields of transplantation, autoimmunity, and tumor immunity.


2003 ◽  
Vol 270 (11) ◽  
pp. 2369-2376 ◽  
Author(s):  
Hazel R. Housden ◽  
Paul J. S. Skipp ◽  
Matthew P. Crump ◽  
Robert J. Broadbridge ◽  
Tom Crabbe ◽  
...  

1996 ◽  
Vol 271 (33) ◽  
pp. 19641-19644 ◽  
Author(s):  
Juliane Bubeck Wardenburg ◽  
Chong Fu ◽  
Janet K. Jackman ◽  
Horst Flotow ◽  
Sandra E. Wilkinson ◽  
...  

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