Related Experiment Video
Updated: Jun 6, 2026
Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays
Published on: September 19, 2018
Comparative analyses of differentially induced T-cell receptor-mediated phosphorylation pathways in T lymphoma cells
Serina Ortiz1, Wenhui Lee, David Smith
1Department of Immunology, Beckman Research Institute, City of Hope, 1450 E. Duarte Road, Duarte, CA 91010-3000, USA.
Abstract:
Activation of T lymphoma cells expressing Syk, but not ZAP-70 tyrosine kinase, has been shown to negatively regulate cell activation and activation-induced cell death (AICD), perhaps due to differential induction of tyrosine phosphorylation modified proteins. To better understand the role of these proteins and their associated molecules/pathways, we studied a previously described model of T lymphoma cells expressing either a kinase-activated chimeric Syk or ZAP-70 genetically linked to T-cell receptor (TCR) ζ chain (Z/Syk or Z/ZAP cells, respectively). To help identify molecules and pathways linked to cell activation or AICD, a comparative semi-quantitative proteomics-based approach was utilized to analyze tyrosine-phosphorylated protein immunoprecipitates from two-minute short-term activated Z/Syk or Z/ZAP cells. Using the resulting bioinformatics data-sets, we identified several differentially immunoprecipitated proteins that could be validated biochemically. More tyrosine-phosphorylated and phosphotyrosine-associated proteins were found in Z/Syk than in Z/ZAP cells. Proteins involved in different unique functional pathways were induced in these cells and showed altered intermolecular interactions in varied pathways. Remarkably, 41% of differentially identified proteins in Z/Syk cells belonged to cell cycle or vesicle/trafficking pathways. In contrast, 21% of such proteins in Z/ZAP cells belonged to metabolism pathways. Therefore, molecular pathways involved in post-translational modifications linked to distinct cellular/physiological functions are differentially activated, which may contribute to varied activation and AICD responses of these cells. In summary, we identified proteins belonging to novel differentially activated pathways involved in TCR-mediated signaling, which may be targets for regulating activation and AICD of T lymphoma cells and for potential cancer therapy.
Insights
This study reveals distinct protein pathways activated by Syk versus ZAP-70 in T lymphoma cells, impacting cell activation and death. These findings offer potential targets for cancer therapy.
Area of Science:
- Immunology
- Molecular Biology
- Proteomics
Background:
- T-cell receptor (TCR) signaling regulates T lymphoma cell activation and activation-induced cell death (AICD).
- Syk and ZAP-70 tyrosine kinases play critical roles in TCR signaling, with differential effects on cell activation and AICD.
- Understanding the downstream molecular pathways influenced by Syk and ZAP-70 is crucial for deciphering T cell responses.
Purpose of the Study:
- To comparatively analyze tyrosine-phosphorylated proteins in T lymphoma cells expressing either Syk or ZAP-70.
- To identify novel molecules and pathways associated with differential T cell activation and AICD.
- To explore potential therapeutic targets for T lymphoma based on TCR-mediated signaling.
Main Methods:
- Utilized a comparative semi-quantitative proteomics approach to analyze tyrosine-phosphorylated proteins.
- Studied T lymphoma cell models expressing kinase-activated chimeric Syk or ZAP-70 genetically linked to the TCR ζ chain (Z/Syk and Z/ZAP cells).
- Performed bioinformatics analysis of proteomics data and biochemical validation of identified proteins.
Main Results:
- Identified significantly more tyrosine-phosphorylated and phosphotyrosine-associated proteins in Z/Syk cells compared to Z/ZAP cells.
- Discovered distinct functional pathways differentially activated in Z/Syk and Z/ZAP cells, with altered intermolecular interactions.
- Found that 41% of differentially identified proteins in Z/Syk cells were involved in cell cycle or vesicle/trafficking, while 21% in Z/ZAP cells were linked to metabolism.
Conclusions:
- TCR-mediated signaling activates distinct molecular pathways involving post-translational modifications, leading to varied T lymphoma cell activation and AICD responses.
- Identified novel pathways and proteins downstream of Syk and ZAP-70 signaling that could serve as targets for regulating T cell function.
- These findings suggest potential therapeutic strategies for T lymphoma and cancer treatment by modulating TCR signaling pathways.
More Related Videos
Related Concept Videos
01:19Interactions Between Signaling Pathways
Convergence and divergence, and cross-talk between signaling pathways
Two distinct signaling pathways can converge on a single functional unit, which may either be a single protein or a complex of proteins. The response is either functionally distinct or synergistic between the two pathways but different from the response...
01:26Receptor Tyrosine Kinases
03:03mTOR Signaling and Cancer Progression
The mTOR pathway or the...