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The Fas-induced apoptosis analyzed by high throughput proteome analysis
C Gerner1, U Frohwein, J Gotzmann
1Institute of Cancer Research, University of Vienna, A-1090 Vienna, Austria. Christopher.Gerner@univie.ac.at
The Journal of Biological Chemistry
|September 9, 2000
Summary
Fas-induced cell death alters cytosolic proteins in Jurkat T-lymphocytes, revealing novel protein synthesis, phosphorylation changes, and degradation, including myosin heavy chain as a caspase target.
Area of Science:
- Cellular Biology
- Proteomics
- Immunology
Background:
- Fas-mediated apoptosis is a critical pathway in T-lymphocyte regulation.
- Understanding proteome dynamics during apoptosis is crucial for deciphering cell death mechanisms.
Purpose of the Study:
- To investigate the proteomic changes in cytosolic proteins during Fas-induced apoptosis in Jurkat T-lymphocytes.
- To identify specific proteins affected by synthesis, degradation, phosphorylation, and localization shifts.
Main Methods:
- Proteome analysis using two-dimensional gel electrophoresis.
- Metabolic labeling and subcellular fractionation for protein synthesis and localization studies.
- Mass spectrometry and Western blotting for protein identification and modification analysis.
Main Results:
- Fas signaling induced synthesis of proteins like hsp27, hsp70B, calmodulin, and H-ras.
- Significant dephosphorylation and phosphorylation events were observed, affecting proteins such as endoplasmin and hsp90.
- Translocation of annexin IV and TCP-1alpha occurred, and myosin heavy chain was identified as a novel caspase target.
- Proteome alterations during Fas-induced apoptosis showed unique characteristics compared to other cell death inducers.
Conclusions:
- Fas-induced apoptosis triggers specific proteomic alterations, including changes in protein synthesis, modification, and localization.
- Myosin heavy chain is a newly identified caspase substrate during apoptosis.
- These proteomic signatures offer insights into the distinct physiological characteristics of various cell death mechanisms.