Document Detail

Alteration of protein composition in mouse thymocytes by signals through T-cell receptor.
MedLine Citation:
PMID:  10870970     Owner:  NLM     Status:  MEDLINE    
To avoid destructive autoimmunity, T-cell precursors (thymocytes) expressing autoreactive T-cell receptor are deleted in the thymus via an apoptotic process by the signals from the T-cell receptor-CD3 complexes. In order to analyze the apoptotic mechanism, we established a cell-free system using the lysates from mouse thymocytes treated in vivo with anti-CD3 monoclonal antibody (mAb). The soluble cytosolic high molecular mass protein fraction from the anti-CD3-treated thymocytes revealed an activity that directly induces nuclear apoptotic morphological changes and DNA fragmentation. This fragmentation activity was not observed in the fraction from the thymocytes without anti-CD3 treatment. Proteins in both fractions were separated by two-dimensional electrophoresis. The silver-stained gels revealed differences in protein spots. These protein spots were identified by database searching of mass spectrometric (MS) and tandem mass spectrometric (MS/MS) data obtained from in-gel tryptic digests of the spots, using an integrated system of liquid chromatography/electrospray ionization/ion-trap mass spectrometry. In this study, the high mobility group protein HMG2 was identified as one of the cytosolic proteins that is increased by the signals from the T-cell receptor, and heterogeneous nuclear ribonucleoprotein A2/B1 and glyceraldehyde 3-phosphate dehydrogenase were found to be decreased by the signals.
T Kawakami; T Nagata; A Muraguchi; T Nishimura
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Electrophoresis     Volume:  21     ISSN:  0173-0835     ISO Abbreviation:  Electrophoresis     Publication Date:  2000 May 
Date Detail:
Created Date:  2000-11-03     Completed Date:  2000-11-03     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8204476     Medline TA:  Electrophoresis     Country:  GERMANY    
Other Details:
Languages:  eng     Pagination:  1846-52     Citation Subset:  IM    
Core Technology Department, Glaxo Wellcome K.K. Tsukuba Research Laboratories, Ibaraki, Japan.
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MeSH Terms
Amino Acid Sequence
Electrophoresis, Gel, Two-Dimensional / methods
Molecular Sequence Data
Proteins / metabolism*
Receptor-CD3 Complex, Antigen, T-Cell / metabolism*
Signal Transduction*
T-Lymphocytes / cytology,  metabolism*
Thymus Gland / cytology
Reg. No./Substance:
0/Proteins; 0/Receptor-CD3 Complex, Antigen, T-Cell

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