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Preparative high-resolution two-dimensional electrophoresis enables the identification of RNA polymerase B transcription factor 3 as an apoptosis-associated protein in the human BL60-2 Burkitt lymphoma cell line

Item Type:Article
Title:Preparative high-resolution two-dimensional electrophoresis enables the identification of RNA polymerase B transcription factor 3 as an apoptosis-associated protein in the human BL60-2 Burkitt lymphoma cell line
Creators Name:Brockstedt, E. and Otto, A. and Rickers, A. and Bommert, K. and Wittmann-Liebold, B.
Abstract:Apoptosis or programmed cell death is essential in the process of controlling lymphocyte growth and selection. We identified RNA polymerase B transcription factor 3 (BTF3), which is associated with anti-IgM antibody-mediated apoptosis, using a subclone of the human Burkitt lymphoma cell line BL60. To identify the transcription factor BTF3, which is expressed only in minor amounts, we used preparative high-resolution two-dimensional gel electrophoresis (2DE) employing carrier ampholytes for isoelectric focusing. Comparison of the 2DE protein patterns from apoptotic and nonapoptotic cells showed BTF3 as a predominantly altered protein spot. The characterization of the differentially expressed transcription factor and 13 marker proteins described in this study were performed by internal Edman microsequencing and/or by peptide mass fingerprinting using matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The proteome analysis was significantly improved by performing the newly developed preparative high-resolution two-dimensional gels employing high protein concentrations.
Keywords:Two-Dimensional Electrophoresis, MALDI-MS, Apoptosis, RNA Polymerase B Transcription Factor 3
Source:Journal of Protein Chemistry
ISSN:0277-8033
Publisher:Kluwer (U.S.A.)
Volume:18
Number:2
Page Range:225-231
Date:February 1999
Official Publication:http://link.springer.com/article/10.1023/A:1020636308270
PubMed:View item in PubMed

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