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Titolo:
ZAP-70 ASSOCIATION WITH T-CELL RECEPTOR ZETA (TCR-ZETA) - FLUORESCENCE IMAGING OF DYNAMIC CHANGES UPON CELLULAR-STIMULATION
Autore:
SLOANLANCASTER J; PRESLEY J; ELLENBERG J; YAMAZAKI T; LIPPINCOTTSCHWARTZ J; SAMELSON LE;
Indirizzi:
NICHHD,CBMB,BLDG 18T,RM 101 BETHESDA MD 20892 NICHHD,SECT LYMPHOCYTE SIGNALING,NIH BETHESDA MD 20892 NICHHD,UNIT ORGANELLE BIOL,CELL BIOL & METAB BRANCH,NIH BETHESDA MD 20892
Titolo Testata:
The Journal of cell biology
fascicolo: 3, volume: 143, anno: 1998,
pagine: 613 - 624
SICI:
0021-9525(1998)143:3<613:ZAWTRZ>2.0.ZU;2-W
Fonte:
ISI
Lingua:
ENG
Soggetto:
PROTEIN-TYROSINE KINASES; TANDEM SH2 DOMAINS; SEVERE COMBINED IMMUNODEFICIENCY; ANTIGEN RECEPTOR; LIVING CELLS; SIGNAL-TRANSDUCTION; ACTIVATION MOTIFS; LYMPHOCYTES-T; BINDING; PHOSPHORYLATION;
Keywords:
ZAP-70; TCR-ZETA; PROTEIN TYROSINE KINASE; INTRACELLULAR SIGNALING; GFP;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
54
Recensione:
Indirizzi per estratti:
Citazione:
J. Sloanlancaster et al., "ZAP-70 ASSOCIATION WITH T-CELL RECEPTOR ZETA (TCR-ZETA) - FLUORESCENCE IMAGING OF DYNAMIC CHANGES UPON CELLULAR-STIMULATION", The Journal of cell biology, 143(3), 1998, pp. 613-624

Abstract

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 TCR zeta chain expressed as a chimeric protein (TT zeta and T zeta zeta), and its mobility in differentintracellular compartments were studied in individual live HeLa cells, using ZAP-70 and T zeta zeta fused to green fluorescent protein (ZAP-70 GFP and T zeta zeta-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 TCR zeta chain is critical for the redistribution, which is enhanced when an active form of the protein tyrosine kinase Lck is coexpressed. Binding specificity to TT zeta was indicated using mutant ZAP-70 GFPs and a truncated zeta chimera. Photobleaching techniques revealed that ZAP-70 GFP has decreased mobility at theplasma membrane, in contrast to its rapid mobility in the cytosol andnucleus. T zeta zeta-GFP is relatively immobile, while peripherally located ZAP-70 in stimulated cells is less mobile than cytosolic ZAP-70in 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 TCR zeta-ZAP-70 interaction and will be a powerful tool for continuing studies of lymphocyte activation.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/11/20 alle ore 19:59:48